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. 2014 Apr 30;20(15-16):2224–2233. doi: 10.1089/ten.tea.2013.0559

FIG. 6.

FIG. 6.

Insulin mediates SOX9 levels and col2a expression. (A) Immunoblot and densitometry show that treatment with HNMPA-(AM3) leads to lower protein levels of Sox6 and Sox9 in P2 grown in SF3D. Each dot represents the mean of an independent experiment; horizontal lines represent mean and 95% CI. n=5 experiments. (B) Gene expression also shows decreases in cartilage-associated genes after downregulating IR. Data are expressed relative to P2-ML and are shown as mean with 95% CI (lower and upper limits). *p≤0.01, **p≤0.05. n=3 experiments. (C) Physical interaction of transcription factor Sox6 and binding protein MED12 with Sox9 is unaffected by HNMPA-(AM3) treatment as seen in co-IP analysis. n=5 independent experiments. (D) Chromatin immunoprecipitation extraction of Sox9-bound DNA showed decreased binding to Col2a1 after HNMPA-(AM3) treatment (IR downregulation). Each dot represents the mean of an independent experiment; horizontal lines represent mean and 95% CI. n=5 experiments. *p≤0.01. C, control; co-IP, co-immunoprecipitation.