FIGURE 7.

UPR activation in L444P GC fibroblasts treated with ERAD inhibitors. Cells were treated with EerI (2 and 6 μm), Kif (50 nm), and MG-132 (0.6 μm) for 24 h. A, Xbp-1 mRNA splicing was determined by RT-PCR followed by gel electrophoresis. B, spliced Xbp-1 band intensities were quantified with National Institutes of Health ImageJ analysis software. Relative mRNA expression levels of CHOP (p < 0.01) (C), ATF4 (p < 0.05) (D), and GC (p < 0.05) (E) were obtained by quantitative RT-PCR and calculated as described in the legend to Fig. 6. The data are reported as mean ± S.D. (error bars). F, Western blot analysis of cells treated with EerI (2 and 6 μm), Kif (50 nm), and MG-132 (0.6 μm) for 48 h using GC specific antibody. GAPDH expression was used as a loading control. G, Western blot band quantification. GC bands were quantified by National Institutes of Health ImageJ analysis software and corrected by GAPDH band intensities.