HSP47 expression protected NIH3T3 cells from Golgi stress-induced apoptosis.

<p>(A) TUNEL-positive cells were significantly increased in HSP47 siRNA-transfected NIH3T3 cells after GalNAc-bn treatment. Results are presented as the percentage of TUNEL-positive cells over total cells, normalized to untreated controls. Data are expressed as the mean (SEM) of at least 3 independent experiments. *p < 0.05 (Student’s <i>t</i> test). (B) The expression of HSP47 or GAPDH protein in NIH3T3 cells after treatment with GalNAc-bn for the indicated periods was examined by western blot analysis using anti-HSP47 or anti-GAPDH antibodies. (C) NIH3T3 cells treated with GalNAc-bn for the indicated periods were examined by western blot analysis using antibody against caspase-2. NIH3T3 cells treated with etoposide (Eto) were used as a positive control for caspase-2 cleavage induction. Cont, untransfected cells; Scr, scrambled siRNA-transfected cells; siRNA, HSP47 siRNA-transfected cells, NoS, no stimulated cells. (D) Western blot analysis showed a remarkable increase in Bcl-xL protein levels in the NIH3T3 cell line stably over-expressing Bcl-xL (BCL) in comparison with NIH3T3 cells expressing the pCAGGS empty vector (CAG). (E) CAG cells and BCL cells treated with GalNAc-bn for 6 h were examined by western blot analysis using an antibody against caspase-2 after HSP47 siRNA transfection.