Fig. 3. Suppression of Beclin expression impairs the autophagy-associated post-translational processing of endogenous LC3-I to LC3-II. (A) Cells were incubated in DMEM + 10% FBS (normal) or HBSS (starved) for 4 hours. (B) Cells were treated with vehicle (–) or 10 µM C₂-ceramide (+) for 24 hours. All cells were subjected to SDS-PAGE and immunoblot analysis to detect the endogenous cytosolic LC3-I and the PE-conjugated LC3-II. The total LC3-II and the ratios of LC3-II to LC3-I represented in the bar graphs were determined from scans of the blots performed with a Kodak 440CF Image Station.