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. Author manuscript; available in PMC: 2010 Feb 1.
Published in final edited form as: Cell Death Differ. 2009 Mar 20;16(8):1135–1145. doi: 10.1038/cdd.2009.28

Figure 6.

Figure 6

Inhibition of autophagy partially rescues the mitochondria loss and apoptosis induced by Bax and PUMA.

A) Saos-2 TetOn Bax cells were transfected with non-targeting siRNA or siRNA against Atg5. 72 hours later, doxycycline was added to induce Bax expression in the presence of z-VAD-fmk. 24 hours after induction, cells were harvested for Western blot analysis. Cell lysates were immunoblotted and probed with antibodies against the indicated proteins. Anti-Atg12 was used to detect Atg5-12 complex to confirm the decrease in Atg5 expression. Anti-COXIV antibody was used to detect mitochondria protein levels.

B) Saos-2 TetOn Bax cells were transfected with non-targeting siRNA as well as siRNA against Atg5, Atg7 and Atg10. 72 hours later, doxycycline was added to induce Bax expression. After 24 hours, cells were harvested and apoptosis was evaluated by flow cytometry. Results are the mean and standard deviation for at least three independent experiments. (* P-value<0.05, **P-value<0.001 compared with non-targeted siRNA transfected cells)

C) Saos-2 TetOn PUMA cells were transfected with non-targeted siRNA or siRNA against Atg5, Atg7 and Atg 10. 72 hours later, doxycycline was added to induce PUMA expression. After 24 hours, cells were harvested and apoptosis was evaluated by flow cytometry. Results are the mean and standard deviation for at least three independent experiments (* P-value<0.05, **P-value<0.001 compared with non-targeted siRNA transfected cells)