Figure 5

Bag5 overexpression modulates the unfolded protein response. A. 22Rv.1 stably overexpressing Bag5 showed increased IRE1/Xbp1 axis activation. Western blot analysis of 22Rv.1 cells stably transfected with pcDNA3.1-HA-Bag5 or the empty vector as control. After treatment with TG for the indicated time points, cells were lysed and subjected to western blot analysis using the indicated antibody. As equal loading control an anti-β-actin antibody was used. B - E. Real time-PCR analysis of 22Rv.1 cells stably expressing the empty vector control (open bars) or HA-Bag5 (filled bars) treated with TG for the indicated time points. Gene expression analysis was performed for CHOP (B), Bcl-2 (C), Bax (D) and the spliced form of Xbp1, Xbp1s (E). Values were normalized to Rib36. Bar charts indicate the mean of at least three independent experiments ± SD (*p <0.05; ** p < 0.01). F. Western blot analysis of 22Rv.1 cells transfected with siRNA targeting GFP or Bag5 and treated with TG for the indicated time points. Anti-α-tubulin was used as equal loading control.