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. 2016 Nov 7;6:36661. doi: 10.1038/srep36661

Figure 4. C/EBPβ reduces adipocytes fatty acid oxidation by negative transcriptional regulation of MC5R.

Figure 4

(A) Fragments of MC5R promoter fused to a luciferase reporter gene were co-transfected into HEK293T cells together with PGL3-basic (control) or pc-C/EBPβ (n = 3). Luciferase activity was corrected for Renilla luciferase activity and normalized to control activity (n = 3). (B) Chromatin immunoprecipitation (ChIP) analysis of MC5R and C/EBPβ (n = 3). (C) Effects of C/EBPβ on FFA level in iWAT adipocytes. Cells were pre-transfected with pc-C/EBPβ or si-MC5R, and then incubated with 500 nM α-MSH for 1 h (n = 3); (D) Effects of C/EBPβ on palmitate oxidation in iWAT adipocytes. Cells were pre-transfected with pc-C/EBPβ or si-MC5R, and then incubated with 500 nM α-MSH for 1 h (n = 3); (E) Effects of C/EBPβ on FFA level in iBAT adipocytes. Cells were pre-transfected with pc-C/EBPβ or si-MC5R, and then incubated with 500 nM α-MSH for 1 h (n = 3); (F) Effects of C/EBPβ on palmitate oxidation in iBAT adipocytes. Cells were pre-transfected with pc-C/EBPβ or si-MC5R, and then incubated with 500 nM α-MSH for 1 h (n = 3); (G) Effects of C/EBPβ on the protein expression of CPT-1, CPT-1 activity and phosphorylation level of ACC in iWAT adipocytes. Cells were pre-transfected with pc-C/EBPβ or si-MC5R, and then incubated with 500 nM α-MSH for 1 h (n = 3); (H) Effects of C/EBPβ on the protein of CPT-1, CPT-1 activity and phosphorylation level of ACC in iBAT adipocytes. Cells were pre-transfected with pc-C/EBPβ or si-MC5R, and then incubated with 500 nM α-MSH for 1 h (n = 3). pc-C/EBPβ: pcDNA3.1-C/EBPβ overexpression vector of C/EBPβ, si-MC5R: lentiviral interference vector of MC5R, CPT-1: carnitine palmitoyl transferase-1, ACC: acetyl-CoA carboxylase, the protein level of CPT-1 was detected by ELISA test. Values are means ± SD. vs. control group, *p < 0.05.