Fig. 3

PPE facilitated autophagosome formation in ASC-differentiated adipocytes (A) Expression levels of LC3A/B in ASC cultured for 8 days with or without PPE (0, 0.1, 0.5 or 1.0 mg/mL) were analyzed using immunoblotting (see Additional Materials and Methods). Experiments were performed in triplicate, and the representative data are shown. Arrowheads indicate LC3A/B-I and LC3A/B-II. The relative intensity of each LC3A/B-II band after normalization to β-actin levels is shown in the lower panel. The blots were cropped, and full-length uncropped blots are presented in Additional Fig. 3. The data are presented as the mean ± standard error of the mean. *p < 0.05 vs. 0 mg/mL PPE. (B) LD and autophagosomes in ASC cultured for 6 days with or without 1.0 mg/mL PPE were stained with Lipi-Blue fluorescent probe and DAPRed, and imaged using a confocal laser scanning microscope. Scale bar: 20 μm. (C) LD and autophagosomes in ASC cultured for 6 days with 1.0 mg/mL PPE were stained with Lipi-Blue fluorescent probe and DAPRed, and imaged using a confocal laser scanning microscope. The images in the lower panel (enlarged) are magnified versions of the inset marked in the upper panel. Scale bar: 50 or 20 μm (enlarged)