Fig. 5

Sox9 phosphorylation at serine 181 is required for maximal transcriptional activation of the Sox10 promoter. a Anti-myc ChIP was performed on SLK^-/- NDL cells transiently transfected with myc-DDK, myc-DDK-Sox9, or myc-DDK-Sox9 S181A. Following anti-myc ChIP, qPCR analysis was performed across two SoxE binding sites within the −6904/−5995 fragment of the Sox10 promoter. qRT-PCR data was normalized to an IgG ChIP (dashed red line). ns: no statistical difference. b Luciferase assays were performed on SLK^-/- NDL cells that were transiently transfected with the indicated plasmids along with the responsive −6904/−5995 element of the Sox10 promoter. Raw light units from the luciferase constructs were normalized to Renilla for each technical sample. Data is represented as the mean luciferase activity from three independent biological replicates +/− SEM. All bars with the same letter are not statistically significant from each other. a compared to b: p < 0.005; a compared to c: p < 0.005; b compared to c: p < 0.05. c SLK^-/- NDL cell were transfected with the indicated plasmids along with the responsive −6904/−5995 element of the Sox10 promoter for 48 h. Cells were treated with heregulin-β (HRG-β), AKT inhibitor (MK-2206), or vehicle control for 2 h prior collecting the cell lysate. Raw light units from the luciferase constructs were normalized to Renilla for each technical sample. Data is represented as the mean luciferase activity from three independent biological replicates +/− SEM. All bars with the same letter are not statistically significant from each other. a compared to b: p<0.05, a compared to c or d: p<0.005, b compared to c: p<0.05, b compared to d: p<0.005 and c compared to d: p<0.05