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. 2012 Sep 11;13(11):1012–1020. doi: 10.1038/embor.2012.133

Figure 4.

Figure 4

Forced HR expression promotes adipogenesis in NIH3T3 and C2C12 cells. (A) Relative expression of Hr in NIH3T3 and 3T3-L1 cells using quantitative reverse transcription–polymerase chain reaction (RT–PCR). (B,C) Analysis of expression of hHR–GFP and GFP alone in NIH3T3 cells (B) and C2C12 cells (C) 24 h after transfection and in non-transfected cells (control), using flow cytometry (upper panels) and fluorescence microscopy (lower panels). Scale bars, 50 μm. Histograms are representative of three flow cytometry analyses. (D,E) Relative expression of adipogenic genes in NIH3T3 cells (D) and C2C12 cells (E) ectopically expressing hHR–GFP or GFP alone prior differentiation, using quantitative RT–PCR. (F) Visualization of lipid incorporation by Oil Red O stains (upper panels) or phase-contrast microscopy (lower panels) in NIH3T3 cells 8 days after differentiation induction. Scale bars, 100 μm. (G) Relative expression of adipogenic genes in NIH3T3 cells on day 8 of differentiation using quantitative RT–PCR. (H) Visualization of lipid incorporation by Oil Red O stains (upper panels) or phase-contrast microscopy (lower panels) in C2C12 cells 8 days after differentiation induction. Scale bars, 100 μm. (I) Relative expression of adipogenic genes in C2C12 cells on day 8 of differentiation using quantitative RT–PCR. (J) Relative expression of myogenic genes in C2C12 cells on day 6 of myogenic differentiation using quantitative RT–PCR. Error bars indicate standard deviations. *P<0.05; **P<0.01; ***P<0.001 (n=3, Student’s t-test). GFP, green fluorescent protein; HR, hairless; PPARγ1, peroxisome proliferator-activated receptor-γ1.