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. 2022 May 25;29(11):2316–2331. doi: 10.1038/s41418-022-01018-8

Fig. 1. Plin2-dependent lipid droplets delay exit from pluripotency.

Fig. 1

a Schematic demonstrating the structure of lipid droplets and the main tissue distribution of perilipin family proteins: WAT white adipose tissue, BAT brown adipose tissue, MSC mesenchymal stem cell. b qRT-PCR analysis of expression of perilipin genes (Plin1, Plin2, Plin3, Plin4, Plin5) in MEF and mESC. Data are mean ± SD, n = 3 biological replicates. Two-tailed unpaired t-tests. ***P < 0.001, **P < 0.005. c qRT-PCR analysis of expression of Plin2, Plin3 and Oct4 during reprogramming of MEFs and spontaneous differentiation of mESCs. Data are mean ± SD, n = 3 biological replicates. d Western blot analysis of Plin2 and Oct4 during reprogramming of MEFs and spontaneous differentiation of mESCs. The experiments were repeated independently three times with similar results. e Immunofluorescence staining of Plin2 (Red) during reprogramming of MEFs and spontaneous differentiation of mESCs. Scale bar, 25 μm. The experiments were repeated independently three times with similar results. f Representative histograms and MFI values of BODIPY 493/503 during reprogramming of MEFs and spontaneous differentiation of mESCs. Data are mean ± SD, n = 3 biological replicates. Two-tailed paired t-tests. ***P < 0.001, **P < 0.005, *P < 0.05. g Schematic depicting experimental design to reduce lipid droplets in mESCs and western blot analysis of Plin2 in WT and Plin2-/- mESCs (KO1 and KO2). The experiments were repeated independently twice with similar results. h Representative images of Oil Red O staining in WT and Plin2-/- mESCs (KO1 and KO2). Scale bar, 100 μm. The experiments were repeated independently three times with similar results. i Triglyceride (TG) content in WT and Plin2-/- mESCs (KO1 and KO2). Data are mean ± SD, n = 3 biological replicates. Two-tailed unpaired t-tests. *P < 0.05. j Representative phase-contrast images and the expression of genes associated with naive pluripotency (Rex1, Tbx3, Klf4) and primed pluripotency (Fgf51, Otx2, Foxa2) of WT and Plin2-/- mESCs (KO1 and KO2) during naive-primed transition on day 5. Scale bar, 100 μm. Data are mean ± SD, n = 3 biological replicates. Two-tailed unpaired t-tests. ***P < 0.001. k Representative phase-contrast and Oct4-GFP images and the expression of pluripotency markers (Oct4, Nanog, Rex1) of WT and Plin2-/- mESCs (KO1 and KO2) on day 3 of differentiation. Scale bar, 100 μm. Data are mean ± SD, n = 3 biological replicates. Two-tailed unpaired t-tests. ***P < 0.001.