Figure 4.

The inhibitory effect of Atg4B mutant on LC3 lipidation is suppressed by exogenous LC3 in a dose-dependent manner. (A) NIH3T3 cells were infected with different amounts of retroviruses bearing mStrawberry-Atg4B^C74A, and stable transformants were selected. The stable cells were cultured in HBSS (Starved) for 1 h, and cell lysates were examined by Western blotting using each antibody. From top panel, anti-Atg4B, anti-RFP, anti-LC3, and anti-α-tubulin. (B) NIH3T3 cells stably expressing mStrawberry-Atg4B^C74A were infected with different amounts of retroviruses bearing GFP-LC3 and then double stable transformants were selected. Parent NIH3T3 cells and the stable transformants were cultured in HBSS (Starved) for 1 h, and cell lysates were examined by Western blotting using each antibody. From top panel, anti-RFP, anti-GFP, anti-LC3, and anti-α-tubulin.