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. 2019 Jan 1;15(1):114–126. doi: 10.7150/ijbs.28679

Figure 3.

Figure 3

COPI mediates reverse transport of EGFRvIII. A: U251EGFRvIII cells were treated with or without 24h serum-free starvation culture, or 24h serum-free starvation culture after adding brefeldin A (BFA) or DMSO for 30min. DMSO is the control of BFA. Immunofluorescence staining were performed to analyze the co-localization of EGFRvIII and calregulin. Right histogram is a comparison of the co-localization coefficients between them. Scale bar, 10μm. B: Immunofluorescence results of the co-localization between EGFRvIII and calregulin in control group and shRNA knockdown group targeting syntaxin6 after treating with or without 24h serum-free starvation culture. Right histogram is a comparison of the co-localization coefficients between them. Scale bar, 25μm. C: U251EGFRvIII cells were treated with or without 24h serum-free starvation culture, or 24h serum-free starvation culture after adding brefeldin A (BFA) or DMSO for 30min, and then immunoblotting analysis were performed to detect the protein levels of nuclear EGFRvIII. DMSO is the control of BFA. Lower histogram was the relative densities of nuclear EGFRvIII. D: U251EGFRvIII cells were treated with or without 24h serum-free starvation culture, or 24h serum-free starvation culture after adding brefeldin A (BFA) or DMSO for 30min, and then immunofluorescence staining were performed to detect the protein levels of nuclear EGFRvIII. DMSO is the control of BFA. Lower histogram was the cell numbers of EGFRvIII's nuclear translocation in 100 cells. Scale bar, 25μm.