Figure 2.
Detachment-induced autophagy in epithelial cells and fibroblasts. (A) Lysates from MCF-10A cells grown attached (A) for 48 h or suspended for the indicated times were immunoblotted with α-LC3 and α-tubulin. (B and C) MCF-10A and 1°HMEC lysates grown attached (A) or suspended for the indicated times were subject to α-LC3 and α-tubulin immunoblotting. When indicated by E/P+, E64d and pepstatin A (10 μg/ml each) were added directly to the culture 4 h before lysis. (D) Cell lysates collected from MDCK2 cells grown attached for 48 h (A) or suspended for the indicated times in complete growth media. When indicated by E/P+, E64d and pepstatin A (10 μg/ml each) were added directly to the culture 4 h before lysis. (E) Wild-type or ATG5−/− MEFs were grown attached for 48 h or detached in complete media for the indicated times. All lysates were subject to immunoblotting with α-LC3 and α-tubulin antibodies.