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. 2016 Mar 10;49(2):207–218. doi: 10.1111/cpr.12246

Figure 2.

Figure 2

Sox9 is a direct target of miR‐30a. (a) The paired miR‐30a seed sequence and the seed‐recognizing site in the wild‐type (WT) and mutant (mut) 3′ UTR of Sox9 are shown as indicated. (b) Chondrocytes were co‐transfected with firefly luciferase reporter plasmids containing either wt‐Sox9 or mut‐Sox9 binding sites and Renilla plasmids (control), negative miRNA control (miR‐NC) or miR‐30a mimics (miR‐30a). The ratio of firefly/Renilla activity was calculated in the cells and normalized to those of the control. The results were expressed as mean ± SD of three independent experiments. *P < 0.05 versus control of wt‐Sox9. (c–f) Chondrocytes were transfected with miR‐30a mimics (miR‐30a) or a negative miRNA control (miR‐NC), and non‐transfected cells were used as a normal control (control). The levels of miR‐30a (c) and Sox9 mRNA (d) were detected by real‐time PCR. Data were normalized to U6 or β‐actin. (e) Sox9 protein levels were detected by western blotting with β‐actin as the loading control. (f) Quantification of Sox9 protein levels was normalized by β‐actin. Data were expressed as mean ± SD of three independent experiments. *P < 0.05 versus Control.