CTX-Fc-BNCs were localized intracellularly at 37°C (Figure 5), which was inhibited by 100nM CPZ, a blocker of clathrin-coated pit formation [21], and by 5mM mβCD (Figure 6), a cholesterol-dislodging oligosaccharide that {Selleck Anti-diabetic Compound Library|Selleck Antidiabetic Compound Library|Selleck Anti-diabetic Compound Library|Selleck Antidiabetic Compound Library|Selleckchem Anti-diabetic Compound Library|Selleckchem Antidiabetic Compound Library|Selleckchem Anti-diabetic Compound Library|Selleckchem Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|Anti-diabetic Compound Library|Antidiabetic Compound Library|buy Anti-diabetic Compound Library|Anti-diabetic Compound Library ic50|Anti-diabetic Compound Library price|Anti-diabetic Compound Library cost|Anti-diabetic Compound Library solubility dmso|Anti-diabetic Compound Library purchase|Anti-diabetic Compound Library manufacturer|Anti-diabetic Compound Library research buy|Anti-diabetic Compound Library order|Anti-diabetic Compound Library mouse|Anti-diabetic Compound Library chemical structure|Anti-diabetic Compound Library mw|Anti-diabetic Compound Library molecular weight|Anti-diabetic Compound Library datasheet|Anti-diabetic Compound Library supplier|Anti-diabetic Compound Library in vitro|Anti-diabetic Compound Library cell line|Anti-diabetic Compound Library concentration|Anti-diabetic Compound Library nmr|Anti-diabetic Compound Library in vivo|Anti-diabetic Compound Library clinical trial|Anti-diabetic Compound Library cell assay|Anti-diabetic Compound Library screening|Anti-diabetic Compound Library high throughput|buy Antidiabetic Compound Library|Antidiabetic Compound Library ic50|Antidiabetic Compound Library price|Antidiabetic Compound Library cost|Antidiabetic Compound Library solubility dmso|Antidiabetic Compound Library purchase|Antidiabetic Compound Library manufacturer|Antidiabetic Compound Library research buy|Antidiabetic Compound Library order|Antidiabetic Compound Library chemical structure|Antidiabetic Compound Library datasheet|Antidiabetic Compound Library supplier|Antidiabetic Compound Library in vitro|Antidiabetic Compound Library cell line|Antidiabetic Compound Library concentration|Antidiabetic Compound Library clinical trial|Antidiabetic Compound Library cell assay|Antidiabetic Compound Library screening|Antidiabetic Compound Library high throughput|Anti-diabetic Compound high throughput screening| inhibits caveolar formation and perturbs clathrin-coated endocytic vesicles [35, 36]. Because 300nM CTX significantly reduced the green fluorescence Inhibitors,research,lifescience,medical of BNCs by competing with CTX-Fc-BNCs (Figure 5(a)), CTX-Fc-BNCs binding on A172
cell surfaces should be specific to the CTX-binding site such as MMP-2 and MT1-MMP. The internalization of CTX-Fc-BNCs was shown to be temperature dependent (Figure 5(b)). This suggests that cellular uptake of CTX-Fc-BNCs was receptor mediated. Zhang et al. reported that CTX was displayed on polyethylene glycol (PEG-) coated iron oxide nanoparticles that were detectable in the tumor lesions of mouse Inhibitors,research,lifescience,medical and rat glioma models. They demonstrated the active targeting of glioma cells using a combination of CTX and supermagnetic or fluorescent compounds in vivo and in vitro [37–40]. CTX-displaying nanoparticles were able to pass the blood-brain barrier (BBB) or the blood-tumor
barrier (BTB) after intravenous injection Inhibitors,research,lifescience,medical and accumulated in brain tumors [38, 41]. Many methods, such as intratumoral injection, intracavity injection, microdialysis, biodegradable polymers, and enhanced convection, have been used for local drug delivery to brain tumors [42]. Given the characteristic features of CTX-Fc-BNCs, the targeted intravenous injection Inhibitors,research,lifescience,medical of brain tumors with nanodrugs displaying CTX-Fcs should alleviate painful side effects in patients. 5. Conclusions We designed a fusion protein between CTX and human IgG-Fcs. Depending on the presence of hinge region of Fc domain, the fusion protein exists as a monomer or
a dimmer. The monomeric form, M-CTX-Fc, performed Inhibitors,research,lifescience,medical as an active targeting ligand to suppress the motility of A172 glioblastoma cells. We then constructed a protein nanocapsule displaying M-CTX-Fc as CTX-Fc-BNCs, which showed specific affinity to the surface of A172 cells and internalized into the cytoplasmic space. This internalization depended on the clathrin-mediated endocytosis pathway. Thus the internalization Histamine H2 receptor was enhanced by the multivalent display of the ligand on nanocapsules, which should be a promising drug delivery system for targeting glioblastoma when an appropriate anticancer agent is loaded. Supplementary Material Figure S1: Confocal microscopic observation for M/D-CTX-Fcs. The M/D-CTX-Fcs attached to cell surfaces at 4°C (upper). One-hour incubation at 37°C promoted the internalization of M/D-CTX-Fcs into cells (lower). The cells were stained with anti-human IgG antibody labeled with FITC. Left: fluorescence image; Right; composite image. M: M-CTX-Fc; D: D-CTX-Fc; Fc: human IgG-Fc. Bars = 10μm. Figure S2: Effect of CPZ on internalization of CTX-Fc-BNCs.