Depletion of endogenous MIF reduces starvation-induced autophagy.

<p>(A) HuH-7-shLuc and HuH-7-shMIF cells were incubated with HBSS with or without NAC for 3-h and 6-h. The cell lysates were collected to detect LC3 conversion by Western blotting analysis. The figure represents one of three independent experiments. (B) HuH-7-shLuc and HuH-7-shMIF cells were incubated in HBSS for starvation. The mRNA changes of autophagy-related genes were analyzed at 0-h, 3-h, and 6-h as indicated. (C) LC3 punctae formation in LC3-EGFP transfected HuH-7-shLuc and HuH-7-shMIF cells 6-h after incubation with HBSS was detected as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0037613#s4" target="_blank">Materials and Methods</a>. (D) The results LC3 punctae formation was calculated by ImageExpress Micro system. **, P<0.01. Data are means ± SD.