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. 2017 Feb 13;8:45. doi: 10.3389/fphys.2017.00045

Figure 1.

Figure 1

Autophagy is necessary for adipocyte differentiation. 3T3-L1 cells were exposed to control medium (FM) or adipocyte differentiation medium (DM) in the presence or absence of the autophagy inhibitors Asn and 3-MA for 14 days. (A) Oil Red O staining shows the effect of autophagy on lipid droplet formation. (B) Quantification of intracellular lipids of 3T3-L1 cells by spectrophotometry. Data represent the mean ± SD. ***P < 0.001 vs. the untreated control (n = 5). (C) Western blot analysis of the expression of the autophagy markers LC3-I, LC3-II and PPARγ with β-actin as the loading control. (D,E) The LC3-II/LC3-I ratio and relative PPARγ expression were quantified by densitometric scanning and graphed. Data indicate the mean ± SD (n = 5). ***P < 0.001 vs. control. (F) Representative fluorescence microscopy images of GFP-LC3 transfected cells treated as indicated. (G) The number of autophagosomes was counted in 10 random fields. Data represent the mean ± SD. ***P < 0.001 vs. the untreated control (n = 10). FM, control medium; DM, differentiation medium.