Fig. 4
siBim attenuates lipoic acid induced-apoptosis 72 h after transfection. a: The cells were treated following protocol of exposure regarding the treatment by lipoic acid (1 mM) administered 24 h after transfection with either 20nM nonspecific siRNA control (siCRTL) or siBim, as described in materials and methods section. b: Bim protein expression level was assessed in control or treated-cells at 72 h post-transfection of IGROV1 (left panel) and IGROV1-R10 (right panel) by western blot. Actin protein is used as a loading control. Actin is a same actin that in Fig. (4e). This blot was performed in the same experiment as that of blot in Fig. 4e. Western blots shown come from one experiment representative of at least three independent experiments and cell lysates. c-d: Morphological features of the cells observed by photon microscopy (left column of each panel) and nuclear features of the cells after DAPI staining (middle column of each panel) were then studied, Bars: 20 μm. DNA content histograms obtained by flow cytometry (right column of each panel) after a 48 h of LA treatment in IGROV1 [c] and IGROV1-R10 (d) cell lines were studied. For each condition, the percentage of sub-G1 and G0-G1 phases is indicated. e: Bcl-xL, Mcl-1, caspases-3 (pro and cleaved forms) protein expression levels were assessed in control or treated-cells at 72 h post-transfection of IGROV1 (left panel) and IGROV1-R10 (right panel) by western blot. Actin protein is used as a loading control. Actin is a same actin that in Fig. (4b). This blot was performed in the same experiment as that of blot in Fig. 4b. Western blots shown are from one experiment representative of at least three independent experiments and cell lysates