Figure 4.
Detachment-induced autophagy in Bcl-2 expressing cells. (A) Punctate GFP-LC3 in detached MCF-10A cells stably expressing GFP-LC3 or coexpressing GFP-LC3 + Bcl-xL suspended for 24 h. Bar, 20 μm. (B) Control (BABE) or Bcl-2–expressing cells were grown attached (A) for 48 h or suspended for the indicated times, lysed, and subject to immunoblotting with α-LC3 and α-tubulin antibodies. (C) Wild-type and Bcl-2–expressing cells were grown attached (A) or suspended for the indicated times, lysed, and subject to immunoblotting with α-catalase and α-tubulin antibodies. (D) Cells transfected with the indicated siRNAs were grown attached (A) or suspended for the indicated times, lysed, and subject to immunoblotting with α-catalase and α-tubulin antibodies. (E) Clonogenic replating efficiency of Bcl-2–expressing cells transfected with the indicated siRNAs after 48 h attachment (white) or suspension (black). Replating efficiency was calculated as described previously. Results are the mean ± SEM from three to five independent experiments.