MET activates autophagy in random skin flaps. (A) Autophagosomes (red) in cells in Area II of flaps in the Control and MET groups by Immunofluorescence staining for LC3II (scale bar, 25μm). (B) IHC for CTSD expression in the ischemic flaps of the Control and MET groups (original magnification × 200; scale bar, 50μm). (C) The optical density values of CTSD were quantified and analyzed in each group. (D&E) Western blotting for Beclin1, LC3II, p62, VPS34, and CTSD expressions in the ischemic flaps of the Control and MET groups. The gels have been run under the same experimental conditions, and cropped blots are used here. (F) Optical density values of Beclin1, LC3II, p62, VPS34, and CTSD were quantified and analyzed in each group. Values are expressed as means± SEM, n= 6 per group. *p< 0.05 and **p< 0.01, vs. Control group.