Figure 2 AFM image and three-dimensional distribution of the MoS 2 film. (a) An AFM image of

the MoS2 nanodisc film deposited on the SiO2/Si substrate. (b) Three-dimensional distribution of the MoS2 nanodiscs. Figure 3a shows XRD patterns of the obtained MoS2 nanodiscs. Because the intensities of the diffraction peaks differed too widely to be presented in a single plot, the larger plot shows the diffraction peaks in the range of 10° to 60°, while the small insert shows the diffraction peaks that appear between 60° and 70°. Over the whole range of diffraction angles, the MoS2 nanodiscs exhibit eight diffraction peaks, located at 14.7°, 29.5°, 33.1°, 47.8°, 54.6°, 56.4°, 61.7°, and 69.2°. They are assigned, respectively, selleck products to the diffraction planes (002), (004), (100), (105), (106), (110), (112), and (108) of MoS2 according to data from the JPDS. The presence selleck kinase inhibitor of these peaks demonstrates that the obtained MoS2 nanodiscs exhibit a variety of crystal structures. Moreover, the obtained diffraction peaks are rather sharp, which shows that the MoS2 nanodiscs are crystalline over a large area. The peak corresponding to the (108) crystal face is much more

intense than the other peaks, indicating that the discs have a strong tendency to adopt the (108) crystal orientation during their growth. Figure 3 Properties of the MoS 2 nanodiscs. (a) XRD pattern of the obtained MoS2 nanodiscs for the diffraction angle in the range of 10° ~ 60°. Inset: the diffraction spectrum of MoS2 nanodiscs for the diffraction angle in the range of 60° ~ 70°. (b) The surface current-voltage curves of the MoS2 nanodiscs. Inset: the layout of four measured points

on the MoS2 disc film. The surface current-voltage (I-V) properties, surface carrier concentration and mobility of the obtained MoS2 nanodiscs are very sensitive to the quality of the film. Figure 3b shows the surface I-V properties of the MoS2 nanodisc film. The inset shows the layout of the four measurement points on the MoS2 Reverse transcriptase nanodisc film. The I-V curves measured between any two points show a perfect linear dependence, which indicates that the deposited MoS2 nanodiscs have good conductivity. The measured carrier concentration of the MoS2 discs is about 3.412 × 106 cm−2, and their electron mobility is as high as 6.42 × 102 cm2/Vs. This mobility value is higher than previously reported values (2 to 3 × 102 cm2/Vs) for single and multilayer MoS2[19, 28]. This significant increase of room-temperature mobility value in our MoS2 may result from the MoS2 nanodisc structure. The mobility of SL MoS2 is generally SCH727965 in vivo smaller than bulk MoS2 because of the larger phonon scattering [29]. However, FL MoS2 exhibits fewer dangling bonds and defect states than does SL MoS2, significantly decreasing the phonon scattering.

Lee JV, Lai S, Exner M, Lenz J, Gaia V, Casati S, Hartemann P, Luck C, Pangon B, Ricci ML, Scaturro M, Fontana S, Sabria M, Sanchez I, Assaf S, Surman-Lee S: An international trial of quantitative PCR for monitoring Legionella in artificial water systems. J Appl Microbiol 2011,110(4):1032–1044. 17. Walker JT, Mackerness CW, Mallon D, Makin T, Williets T, Keevil CW: Control of Legionella pneumophila in a hospital water system by chlorine dioxide. J Ind Microbiol 1995,15(4):384–390.PubMedCrossRef GS1101 18. Yanez MA, Nocker A, Soria-Soria E, Murtula R, Martinez

L, Catalan V: Quantification of viable Legionella pneumophila cells using propidium monoazide combined with quantitative PCR. J Microbiol Methods 2011,85(2):124–130.PubMedCrossRef 19. Nogva HK, Drømtorp SM, Nissen H, Rudi K: Ethidium monoazide for DNA-based differentiation of viable and dead bacteria by 5′nuclease PCR. Biotechniques 2003,34(4):804–808.PubMed Competing interests The authors declare that they have no competing interests. Authors’ contributions LHK: Was involved in the decision LY333531 price making on choosing the locality of apartments and tap locations. Collected most samples. Helped during concentration and cultivation of samples. Purified

DNA from samples and tested them on qPCR. Had the main responsibility and workload of all data analyses. Was active in the interpretation of the results. Wrote the article. Has read and approved the final manuscript. SU: Has participated actively in all parts Sodium butyrate of the process. Responsible for the diagnostics of patients and environmental isolates together with unravelling the source of infection. Responsible for the culture analysis of water samples. Involved in AZD5363 order decisions about choosing the locality of apartments and tap locations. Involved in preparation

of the manuscript. Has read and approved the final manuscript. KAK: Contributed to designing the study, involved in discussing the results and building the article. Has read and approved the final manuscript. HANA: Contributed to planning, judging and interpretation of the results and building the article. Has read and approved the final manuscript.”
“Background The marine green alga Bryopsis has long been suspected to harbor endogenous bacteria. These intracellular bacteria have been repeatedly observed in the cytoplasm as well as vacuolar regions of algal thalli and gametes by electron microscopy [[1, 2] and personal observations see additional file 1], suggesting the presence of bacterial endophytes within Bryopsis is a natural phenomenon. Recently, the first insights were provided into the identity and diversity of these bacterial endophytes within two Bryopsis species from the Pacific Mexican coast [3]. Full length 16S rRNA gene analysis showed that the Bryopsis endophytic bacterial communities are quite low in diversity (i.e.

Rarefaction analysis Rarefaction analysis at the most resolved level of the NCBI taxonomy in MEGAN showed the taxonomic richness detected in the sediment samples (Figure 2). Including all PX-478 research buy assigned taxa, 1034 and 882 leaves were detected in the 0-4 cm and 10-15 cm metagenome respectively. Of these, 785 (0-4 cm) and 596 (10-15 cm) were bacterial Epigenetics inhibitor and 58 (0-4 cm) and 127 (10-15 cm) archaeal. The rarefaction curves for bacterial and total taxa indicated that not all the taxonomic richness in the sediment was accounted for in our metagenomes. Still, the curves were levelling off from a straight line already at 10% of the metagenome

size indicating repeated sampling of Selleck VX809 the same taxon. It is therefore likely that abundant taxa in the sediments were accounted for in the two metagenomes. Figure 2 Rarefaction curves created in MEGAN. Rarefaction analysis was performed at the most resolved taxonomic level of the NCBI taxonomy in MEGAN for each metagenome. The curves for all taxa include Bacteria, Archaea, Eukaryota, Viruses, unclassified and other sequences.

While most of the archaeal taxa in the 10-15 cm metagenome were accounted for, the number of taxa in the 0-4 cm was still increasing at 100% sampling. This difference is likely due to the low abundance of Archaea in the 0-4 cm metagenome (0.97% of reads) compared to the 10-15 cm metagenome (18.09% of reads) as shown in Figure 3. Figure 3 Normalized MEGAN tree at the domain level. Comparative tree view of the two metagenomes from the root to the domain level. The 0-4 cm metagenome

is presented in red and the 10-15 cm metagenome in blue. The numbers in brackets give the percentage of total reads assigned to each node for the two metagenomes. The size of the individual nodes is scaled logarithmically to indicate number of reads assigned. Taxonomic binning There was a significant difference in the proportion of reads assigned to Bacteria and Archaea for the two metagenomes (Figure 3). In the 0-4 cm metagenome 60.87% of the reads were assigned to Bacteria 5-Fluoracil research buy and 0.97% to Archaea, while in the 10-15 cm metagenome 47.14% of the reads were assigned to Bacteria and as much as 18.09% to Archaea. This shift in the prokaryotic community structure suggests that Archaea thrive better and thereby also are likely to contribute more to the metabolism in the 10-15 cm sediment horizon. Xipe analyses of the binned reads (confidence cut-off of 0.95, 0.98 and 0.99) at the phylum level (Table 1) and at the genus level (Additional file 2, Tables S2 and Additional file 3, Table S3) showed a significant difference between the two metagenomes as to the most abundant taxa [25]. The high abundance of Archaea in the 10-15 cm metagenome compared to the 0-4 cm metagenome was striking at the phylum level as well (Table 1).

The increase in overall average peak and mean power with BTE supplementation implies increased performance with BTE which may be due to increased recovery between intervals of the WAnT protocol. As well, the blood lactate levels were higher with BTE supplementation at 0 and 5 min post high intensity exercise which is consistent with the higher workload completed. Based on these power and lactate results, the BTE supplementation AZD6244 in vivo appears to have resulted in the performance of more total work, which amplifies the significance of the biochemical and hormonal findings. BTE supplementation,

therefore, may not have only sped the recovery from the oxidative stress response, but may have also blunted the response as the anticipated increase in appearance of oxidative stress and inflammatory markers with increase in workload was not observed. Future research on BTE supplementation should focus on the link between the acute physiological effects and the long-term outcome of increased anaerobic exercise performance over a longer duration of time. Evaluating the effect of BTE supplementation on the performance of progressive anaerobic exercise training would aid in elucidating the pathway from reduced oxidative stress, HPA recovery, and DOMS responses to increased performance Fosbretabulin molecular weight and enhanced fitness. Inflammation, oxidative stress, and the occurrence of DOMS following

high intensity anaerobic exercise are essential processes for acquisition of strength and LGX818 nmr muscle hypertrophy after exercise [1, 10, 12]. In excess, these responses delay recovery and result in reduced power and performance. In theory, improved training and performance would result from reducing the length of recovery and/or the extent of muscle damage after a high-intensity exercise bout. Theaflavins, found in black tea extract, have been observed Megestrol Acetate to have antioxidant effects [4] as well as anti-inflammatory effects [8, 19]. Multiple epidemiological studies have found an inverse association with black tea consumption and chronic disease incidence/mortality including: congestive heart disease, stroke, atherosclerosis, pancreatic,

bladder, and prostate cancers [7]. These findings have led to numerous studies examining the antioxidant effects of tea polyphenols, with emphasis on green tea and its catechins, in several models of disease. The use of antioxidants to improve exercise performance and reduce muscle soreness is not a new concept. In this capacity, the antioxidant effects of vitamin C and E have been extensively researched [16–18]. Green tea extract (GTE) and its effects on exercise capacity and metabolism have been examined in mouse models. The duration of treadmill running was prolonged in BALB/c mice given GTE [23]. Exercise combined with GTE had a synergistic effect in attenuating high fat diet induced obesity in C57BL/6J mice [24].

Mol Cancer Ther 2009, 8:2375–2382.PubMedCrossRef 25. Li H, Simpson ER, Liu JP: Oestrogen, telomerase, ovarian ageing and cancer. Clin Exp Pharmacol Physiol 2010, 37:78–82.PubMedCrossRef 26. Spinella F, Rosano L, Del DM, Di C, Nicotra MR, Natali PG, Bagnato A: Endothelin-1 inhibits prolyl hydroxylase domain 2 to activate Nepicastat in vitro hypoxia-inducible JPH203 order factor-1alpha in melanoma cells. PLoS One 2010, 5:e11241.PubMedCrossRef 27. Goteri G, Lucarini G, Zizzi A, Rubini C, Di PR, Tranquilli AL, Ciavattini A: Proangiogenetic molecules, hypoxia-inducible

factor-1alpha and nitric oxide synthase isoforms in ovarian endometriotic cysts. Virchows Arch 2010, 456:703–710.PubMedCrossRef 28. Knechtel G, Szkandera J, Stotz M, Hofmann G, Langsenlehner U, Krippl P, Samonigg H, Renner W, Langner C, Dehchamani D, Gerger A: Single nucleotide polymorphisms in the hypoxia-inducible factor-1 gene and colorectal cancer risk. Mol Carcinog 2010, 49:805–809.PubMed 29. Miyazawa M, Yasuda M, Fujita M, Hirabayashi K, Hirasawa T, Kajiwara H, Muranmatsu T, Miyazaki S, Harasawa M, Matsui N, Ogane N, Murakami M, Mikami M, Yanase T, Osamura RY: Granulosa cell tumor with

activated mTOR-HIF-1alpha-VEGF MAPK inhibitor pathway. J Obstet Gynaecol Res 2010, 36:448–453.PubMedCrossRef 30. Villaume K, Blanc M, Gouysse G, Walter T, Couderc C, Nejjari M, Vercherat C, Cordire-Bussat M, Roche C, Scoazec JY: VEGF secretion by neuroendocrine tumor cells is inhibited by octreotide and by inhibitors of the PI3K/AKT/mTOR pathway. Neuroendocrinology 2010, 91:268–278.PubMedCrossRef 31. Zeng M, Kikuchi H, Pino

MS, Chung DC: Hypoxia activates the K-ras proto-oncogene to stimulate angiogenesis and inhibit apoptosis in colon cancer cells. PLoS One 2010, 5:e10966.PubMedCrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions PZ carried out the proliferation, cell cycle and apoptosis assay, participated in drafted the manuscript. YN carried out the invasion experiment, participated in experiment design and drafted the manuscript. LY conceived of the study, participated in its design and coordination, performed the statistical analysis and helped to draft the manuscript. MC carried out the telomerase activity assay, participated in the draft Methamphetamine preparation. CX participated in the design of the study and performed the statistical analysis. All authors read and approved the final manuscript. Authors’ informations PZ, M.D., medical master candidate, Dept. Gynecology, Obstetrics & Gynecology Hospital, Fudan University; senior medical registrar, Dept. Obstetric & Gynecology, Shangyu City Hospital; YN, M.D. & Ph.D., assistant professor, Dept. Physiology & Pathophysiology, Shanghai Medical College, Fudan University; LY, M.D. & Ph.D., associate professor & medical consultant, Dept. Gynecology, Obstetrics & Gynecology Hospital, Fudan University; MC, M.B., medical master candidate, Dept.

Further, several investigators report that SpiC is required for the translocation of SPI-2 effector proteins into the target cells by interacting with SsaM,

a SPI-2 encoded protein [10–12]. In addition to these reports, we have shown that SpiC contributes to Salmonella-induced activation of the signal transduction pathways in macrophages, leading to the production of mediators such as interleukin-10, prostaglandin E2, and the expression of the suppressor in cytokine signaling 3 (SOCS-3) that are thought to have important roles in Salmonella virulence [13–15]. Additionally, our recent study shows that SpiC is involved in the expression of FliC, a component of the flagella filaments, where FliC plays a significant role in SpiC-dependent activation of the signal transduction check details pathways Selleck Crenigacestat in macrophages

following Salmonella infection [16]. However, the mechanism of how SpiC affects the expression of FliC remains unknown. The flagellum is essential for bacterial motility. Its structure consists of a basal body, a hook, and a filament. In Salmonella, synthesis of the flagellum involves over 50 genes. The expression of these genes is organized into three hierarchies. At the top hierarchy is the class 1 flhDC operon and it is essential for transcription of all of the genes for the flagellar cascade. flhDC expression is influenced at the transcription or post-transcription level by a number of global regulatory factors. The class 2 operons contain genes encoding the hook-basal body-associated proteins, a few regulatory proteins, and a component of the flagellum-specific type III Thiazovivin cost export pathway. The class 3 operons contain genes involved in filament formation, flagella rotation and chemotaxis [17, 18]. Flagellin,

a component of the filament, is transported from the cytoplasm using the flagellum-specific type III export system in the basal body where it is polymerized with the help of the cap protein FliD [19, 20]. This results in the assembly of the long helical flagella filaments. S. enterica serovar Typhimurium expresses two antigenically distinct flagellins encoded by the fliC and fljB genes and are coordinately expressed using a phase-variation mechanism [17]. FliC also has a role Reverse transcriptase as a potent stimulator of the immune and pro-inflammatory responses [21, 22]. Several reports show that FliC activates the signal transduction pathways via Toll-like receptor 5 (TLR5) in cultured cells (e.g. epithelial cells) leading to the induction of immune and pro-inflammatory genes [23–26]. In addition to TLR5, flagellin was recently shown to be recognized in the host cell cytosol by two different Nod (nucleotide-binding oligomerization domain)-like receptors, Ipaf and Naip5 (also known as Birc1e) [27, 28]. Here, we investigate the mechanism of how SpiC regulates flagellum synthesis in S. enterica serovar Typhimurium.

However, prospective studies with larger populations are required to determine whether S. tigurinus is a commensal or an opportunistic oral pathogen with a potential for development of invasive infections. Acknowledgment The study was supported by the University of Zurich. We thank the laboratory technicians for their dedicated help. References 1. Marsh PD: Are dental diseases examples of ecological catastrophes?

Microbiology 2003, 149(Pt 2):279–294.PubMedCrossRef 2. Albandar JM: Underestimation of periodontitis in NHANES surveys. J Periodontol 2011, 82(3):337–341.PubMedCrossRef 3. Konig J, Holtfreter B, Kocher T: Periodontal health in Europe: future trends based on treatment needs learn more and the provision of periodontal services–position paper 1. Eur J Dent Educ 2010, 14(Suppl 1):4–24.PubMedCrossRef 4. Marcenes W, Kassebaum NJ, Bernabe E, Flaxman A, Naghavi M, Lopez A, Murray CJ: Global burden of oral conditions in 1990–2010: a systematic analysis. J Dent Res 2013, 92(7):592–597.PubMedCrossRef 5. Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE: Defining the normal bacterial flora of the oral cavity. J Clin Microbiol 2005, 43(11):5721–5732.PubMedCentralPubMedCrossRef 6. Papapanou PN, Behle JH, Kebschull M, Celenti R, Wolf DL, Handfield M, Pavlidis P, Demmer RT: Subgingival bacterial colonization profiles correlate with gingival

VX-680 supplier tissue gene expression. BMC Microbiol 2009, 9:221.PubMedCentralPubMedCrossRef 7. Socransky SS, Haffajee AD: Implications of periodontal microbiology for the treatment of periodontal infections. Compt Rendus Geosci 1994, 18:S684–S685. 688–693; quiz S714-687. 8. Lalla E, Papapanou PN: Diabetes mellitus and periodontitis: a tale of two common interrelated diseases. Nat Rev Florfenicol Endocrinol 2011, 7(12):738–748.PubMedCrossRef 9. Beck JD, Offenbacher S: Systemic effects of periodontitis: epidemiology of periodontal disease and cardiovascular disease. J Periodontol 2005, 76(11 Suppl):2089–2100.PubMedCrossRef 10. Spellerberg B, Brandt C: Streptococcus. In Manual of clinical microbiology.

Volume 1. 10th edition. Edited by Versalovic J, Carroll KC, Funke G, Jorgensen JH, Landry ML, Warnock DW. Washington, DC: ASM Press; 2011:331–349. 11. Zbinden A, Mueller NJ, Tarr PE, Sproer C, Keller PM, Bloemberg G: Streptococcus tigurinus sp. nov., isolated from blood of patients with endocarditis, meningitis and spondylodiscitis. Int J Syst Evol Microbiol 2012, 62(Pt 12):2941–2945. 12. Zbinden A, Mueller NJ, Tarr PE, Eich G, Schulthess B, Bahlmann AS, Keller PM, Bloemberg GV: Streptococcus tigurinus , a novel member of the Streptococcus mitis group, Selleckchem MRT67307 causes invasive infections. J Clin Microbiol 2012, 50(9):2969–2973. 13. Zbinden A, Quiblier C, Hernandez D, Herzog K, Bodler P, Senn MM, Gizard Y, Schrenzel J, François P: Characterization of Streptococcus tigurinus small-colony variants causing prosthetic joint infection by comparative whole-genome analyses. J Clin Microbiol 2014, 52(2):467–474. 14.

: Causes of bovine abortion, stillbirth and neonatal death in Finland 1999–2006. Acta Vet Scan 2007, 49:S3.CrossRef 30. Santini F, Borghetti V, Amalfitano G, Mazzucco A: Bacillus licheniformis prosthetic aortic-valve endocarditis. J Clin Microbiol 1995, 33:3070–3073.PubMed 31. Tabbara KF, Tarabay N: Bacillus licheniformis corneal ulcer. Am J Ophthalmol 1979, 87:717–719.PubMed 32. Sugar AM, Mccloskey RV: Bacillus licheniformis sepsis. J Am Med Assoc 1977, 238:1180–1181.CrossRef 33. Kramer JM, Gilbert

ATM Kinase Inhibitor supplier RJ: Bacillus cereus and other Bacillus species. In Foodborne bacterial pathogens. Edited by: Doyle MP. New York: Marcel Dekker Inc; 1989:21–70. 34. Salkinoja-Salonen MS, Vuorio R, Andersson MA, Kampfer P, Andersson MC, Honkanen-Buzalski T, et al.: Toxigenic strains of Bacillus licheniformis related to food poisoning. Appl Environ Microbiol 1999, 65:4637–4645.PubMed 35. Mikkola R, Kolari M, Andersson MA, Helin J, Salkinoja-Salonen MS: Toxic lactonic lipopeptide from food poisoning isolates of Bacillus licheniformis . Eur J Biochem 2000, 267:4068–4074.PubMedCrossRef 36. Errington J: Regulation of endospore formation in Bacillus subtilis . Nature Rev Microbiol 2003, 1:117–126.CrossRef 37. Setlow P: Spores of Bacillus subtilis : their resistance to selleck and killing by radiation, heat and chemicals. J Appl Microbiol

2006, 101:514–525.PubMedCrossRef 38. Setlow P, Johnson EA: Spores and their significance. In Food microbiology: fundamentals and frontiers. Edited by: Doyle MP, Beuchat LR. Washington, DC: ASM Press; 2007:35–67. 39. Zuberi AR, check details Feavers IM, Moir A: Identification of 3 complementation units in the gerA spore germination locus of Bacillus subtilis . J Bact 1985, 162:756–762.PubMed 40. Feavers IM, Miles JS, Moir A: The nucleotide sequence Clomifene of a spore germination gene ( gerA ) of Bacillus subtilis 168. Gene 1985, 38:95–102.PubMedCrossRef 41. Zuberi AR, Moir A, Feavers IM: The nucleotide-sequence and gene organization of the gerA spore germination operon of Bacillus subtilis 168. Gene 1987, 51:1–11.PubMedCrossRef 42. van der Voort M, Garcia D, Moezelaar R, Abee T: Germinant receptor diversity and germination responses of four strains

of the Bacillus cereus group. Int J Food Microbiol 2010, 139:108–115.PubMedCrossRef 43. Paredes-Sabja D, Setlow P, Sarker MR: Germination of spores of Bacillales and Clostridiales species: mechanisms and proteins involved. Trends Microbiol 2011, 19:85–94.PubMedCrossRef 44. Ross CA, Abel-Santos E: Guidelines for nomenclature assignment of Ger receptors. Res Microbiol 2010, 161:830–837.PubMedCrossRef 45. Halmann M, Keynan A: Stages in germination of spores of Bacillus licheniformis . J Bact 1962, 84:1187–1193.PubMed 46. Martin JH, Harper WJ: Germination response of Bacillus licheniformis spores to amino acids. J Dairy Sci 1963, 46:663–667.CrossRef 47. White CH, Chang RR, Martin JH, Loewenst M: Factors affecting L – Alanine induced germination of Bacillus spores. J Dairy Sci 1974, 57:1309–1314.PubMedCrossRef 48.

) under the luminescence setting. Viability at each motesanib or imatinib concentration was expressed as a percentage of the vehicle control (0.2% DMSO). Results In Vitro Inhibition of Wild-Type Kit by Motesanib Motesanib potently inhibited SCF-find more induced autophosphorylation of Kit in CHO cells stably transfected with the wild-type KIT gene (IC50 = 36 nM). In comparison,

imatinib inhibited wild-type Kit with an IC50 of 165 nM. Inhibition of Wild-Type Kit Activity in Mice by Motesanib Hair depigmentation was used as a surrogate marker to assess the ability of motesanib to inhibit Kit activity in vivo [16]. Following depilation, female C57B6 mice were administered either 75 mg/kg motesanib (n = 8) or vehicle (n = 8) twice daily for 21 days. In mice receiving motesanib, hair regrowth was markedly depigmented compared with mice receiving SN-38 chemical structure Akt inhibitor drugs vehicle (Figure 1). This effect was reversible. Following the cessation of motesanib treatment on day 21, the mice were depilated again on day 28. There was no apparent depigmentation of regrown hair on day 35. Similar results were obtained in male mice (data not shown). Figure 1 Effect of treatment with motesanib or vehicle on hair depigmentation, a surrogate marker of Kit activity [16], in female C57B6 mice. Anesthetized animals were depilated and immediately treated with

either vehicle (water; left panels) or motesanib 75 mg/kg BID (right panels) for 21 days. On day 21, hair depigmentation was assessed. Depilation was repeated on day 28 and hair depigmentation was again assessed on day 35. Representative images from each treatment group for the day-21 and day-35 time points are shown. BID = twice daily. Characterization of Kit Mutants Figure 2 summarizes the results from the autophosphorylation experiments using CHO cells stably transfected with the wild-type KIT gene or various KIT mutant genes. Tyrosine phosphorylation of wild-type Kit was

dose-dependent, with the greatest intensity of autophosphorylation occurring after a 30 minute incubation of the cells with 300 ng/mL of SCF. In contrast, tyrosine phosphorylation of activated Etomidate Kit mutants occurred in the absence of SCF with no further phosphorylation induced by treatment with SCF. Figure 2 Effect of stem cell factor (SCF) treatment on tyrosine phosphorylation of wild-type Kit and mutant Kit isoforms stably expressed in Chinese hamster ovary cells. Chinese hamster ovary cells stably transfected with wild-type (WT) or mutant KIT isoforms were stimulated with single serial dilutions of stem cell factor, and Kit phosphorylation was assessed. For mutant Kit isoforms, data are expressed as the percentage of vehicle control. For wild-type Kit, data are expressed as the percentage of phosphorylation observed following stimulation with 300 ng/mL SCF. The results of a single experiment are shown.