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. Author manuscript; available in PMC: 2010 Mar 1.
Published in final edited form as: Cell Metab. 2009 Mar;9(3):287–302. doi: 10.1016/j.cmet.2009.01.013

Figure 4.

Figure 4

Sox9 directly binds C/EBPβ and C/EBPδ promoters, and suppresses their transcription. (A) RT-qPCR for various transcription factors 72 h after Sox9 transfection into 3T3-L1 cells. *, P<0.05 and **, P<0.01 compared to cells transfected with empty vector. (B) C/EBPb and C/EBPd promoter activity after co-transfection with Sox9 into 293FT cells. pRL-SV40 was used as internal control. Results are mean ± SEM; **, P<0.01 (C) Numbering denotes primer sets in the C/EBPβ and C/EBPδ promoter regions used for ChIP (left panel). ChIP for C/EBPβ and C/EBPδ using 3T3-L1 cells (right panel). Bar; sites with core consensus Sox9 binding sequence, Star; actual Sox9 binding sites. (D) EMSA for Sox9 binding sites. In vitro translated Sox9 protein and 32P labeled oligonucleotides were used for EMSA. Probe only (lane 1); 1 μl (lane 2) and 2 μl (lane 3) of Sox9 translation reaction with probe; 1 μl of Sox9 with 100 × wild-type competitor (lane 4); 1 μl of translation reaction of empty pcDNA3.1 vector with probe (lane 5); 1 μl of Sox9 with 100 × mutant competitor (lane 6).