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. 2014 Jun 11;5(13):5138–5152. doi: 10.18632/oncotarget.2094

Figure 3. Bak and Bax-dependent apoptosis by ERBB3 knockdown in HCT116 cells.

Figure 3

(A) Cell proliferation and (B) cell cycle distribution relative to siCTRL (–) were determined 72 h after treatment with siERBB3 (+) in HCT116 mutant cells. Numbers in open box indicate a percent of G1 populations. (C) Protein extracts were prepared 72 h after siERBB3 treatment and analyzed by western blotting with indicated antibodies (left). Parp1 cleavage was quantified (right). (D) Protein extracts in the cytosol or membrane/organelle fraction 48 h after siERBB3 treatment were analyzed by western blotting in Bak-/- cells (top). The relative band intensities of mitochondrial Bax to cytosolic Bax were normalized to that of siCTRL (bottom). (E) Cell proliferation, (F) FACS and (G) western blotting (left) and quantification of parp1 cleavage (right) were performed after serial treatments with siBak and siERBB3. The cells were analyzed 48 h after the second siRNA treatment. For the knockdown, 5 nM of siERBB3 or 0.2 nM of siBak were used. siERBB3 group was statistically compared to siCTRL group at each point, unless otherwise indicated.