Regulation of Autophagy-Related Protein and Cell Differentiation by High Mobility Group Box 1 Protein in Adipocytes

Figure 4

Effects of recombinant HMGB1 protein on autophagy in various differentiation periods of 3T3-L1 adipocytes. Differentiation of 3T3-L1 preadipocytes was induced at 0, 2, 4, 6, 8, and 10 days. The adipocytes were added with 0.2 μg/mL recombinant HMGB1 protein 24 h before differentiation. (a) Pictures were taken with a bright-field microscopic images of differentiating cells. (b) Oil-Red-O staining (200 magnification). (c) The expression of proteins HMGB1, LC3, and P62 was detected by Western blot. (d–f) HMGB1, LC3, and P62 protein were quantified and normalized to GAPDH. (g-h) Levels of PPAR-γ and FABP4 were determined by quantitative PCR in differentiation at 0, 4, and 8 days, , , , and versus the day itself (control).

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