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

Figure 1.

Figure 1

Hairless is required for adipocyte differentiation in vitro. (A) Relative hairless (HR) expression in 3T3-L1 cells at indicated time points of differentiation using quantitative reverse transcription–polymerase chain reaction (RT–PCR). (B) HR expression in 3T3-L1 cells at indicated time points of differentiation using western blotting. GCN5 was used as a loading control for nuclear extracts. (C) Determination of efficiency of HR knockdown in non-stimulated HR-depleted (shHr1 and shHr2) and non-silenced (ns) 3T3-L1 cells using quantitative RT–PCR. (D) Visualization of lipid incorporation by Oil Red O stains (left panels) and phase-contrast microscopy (right panels) in 3T3-L1 cells 8 days after differentiation induction. Scale bars, 50 μm. (E) Relative expression of CCAAT/enhancer-binding protein-β (C/EBPβ) and C/EBPδ in 3T3-L1 cells before (0 h) and 8 h after stimulation of adipogenesis using quantitative RT–PCR. (F) Relative expression of peroxisome proliferator-activated receptor-γ1 (PPARγ1) and PPARγ2 in 3T3-L1 cells at indicated time points of differentiation using quantitative RT–PCR. (G) Relative expression of C/EBPα, fatty acid binding protein 4 (FABP4) and adiponectin in 3T3-L1 cells before (0 h) and 8 days after stimulation of adipogenesis using quantitative RT–PCR. (H) Expression of HR, PPARγ1, PPARγ2, C/EBPα, fatty acid synthase (FASN) and FABP4 in 3T3 cells at indicated time points of differentiation using western blotting. The histone acetyl transferase GCN5 was used as a loading control for nuclear extracts. Tubulin was used as a loading control for total extracts. (I) Determination of efficiency of HR knockdown in non-stimulated HR-depleted (shHr1 and shHr2) or non-silenced (ns) primary mouse preadipocytes using quantitative RT–PCR. (J) Visualization of lipid incorporation by phase-contrast microscopy in primary mouse preadipocytes 8 days after differentiation induction. Scale bars, 50 μm. Error bars indicate standard deviations. *P<0.05; **P<0.01; ***P<0.001 (n=3 for B,C,E,F,G,I; n=4 for A, Student’s t-test). NS, nonsignificant; sh, short hairpin.