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. 2019 Dec 13;10:841. doi: 10.3389/fendo.2019.00841

Figure 4.

Figure 4

Leptin recovers 3T3-L1 differentiation in the absence of insulin. 3T3-L1 adipocytes were cultured with no hormone, insulin or leptin (4 or 40 nM) up to 3, 10, or 17 days after induction of differentiation. (A) Fluorescence images of cells stained with Bodipy (green) for lipid droplets and DAPI (blue) for nuclei. Images are representative of 3–5 independent experiments. Scale bar represents 50 μm. (B) Quantification of the green area per cell in each field. Data are depicted as the fold change relative to the group with no hormone supplementation. Bars represent mean ± standard error of the mean of 3–5 independent experiments. (C) Western blot analysis of PLIN1, CAV-1, SREBP1C, PPARγ, and β-actin. Blots are representative of three independent experiments in the cases of PLIN1, CAV-1, and PPARγ and two independent experiments in the case of SREBP1C. Cell lysates were analyzed in the same gel and cropped for clearer comparison among groups. *p < 0.05 relative to the group in which cells were cultured with neither insulin nor leptin. Statistical analyzes were performed by using the Kolmogorov-Smirnov test.