FIGURE 5.

Overexpression of miR-675 up-regulates COL2A1 expression and rescues COL2A1 levels in H19- and SOX9-depleted HACs. Overexpression of miR-675 (10 nm) significantly up-regulates COL2A1 mRNA (A) (*, p < 0.05) and secreted protein levels (B) in HACs (1 nm and 10 nm of miR-675 used). H19 depletion resulted in reduced levels of secreted COL2A1, whereas SOX9 was unaffected (C). In addition, overexpression of miR-675 (10 nm) rescues COL2A1 levels in H19- and SOX9-depleted HACs (D). Expression of COL9A1 was not affected by depletion of H19 or by overexpression of H19-derived miR-675 (E). In all experiments, HACs were incubated in 1% oxygen for 3 days before analysis. p, passage number. Gene expression is relative to that of ribosomal protein RPLP0. Ctrl, negative control siRNA-targeting luciferase; si, small interfering.