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(A) Baseline expression of miR-204 and miR-211 in undifferentiated C3, C6, C12, BM12 MSCs by Q Real Time PCR. Full scale bar graph. (B) Modulation of RUNX-2 expression. miR-204 and miR-211 overexpression inhibit RUNX-2 as evaluated by Western blot in C3 MSCs 1wk chondroid cultures. CRTAP, a cartilage master gene and a potential target of miR-204&211 according to Target Scan6.2 (www.targetscan.org), but not miR and a (www.microrna.org), is not affected in miR 204 and miR 211 overexpressing C3 MSCs, whereas RUNX-2 is significantly decreased. GAPDH is the control protein and is steady. (C) RUNX-2 protein expression level evaluated in control C3, C3 miR204 and C3 miR211 compared with wild type C6 and C13 primary human colonic MSCs. Actin is used as control protein. (D) Modula4tion of RUNX-2 protein expression in wild type C3 MSCs upon induction of chondroid differentiation (a) or C3 over-expressing miR-204&211 (b). RUNX-2 was down-regulated during ex vivo chondrogenesis (a) Analyses were conducted on cartilage pellets. MiR-204&211 strongly down-regulated RUNX-2 protein expression in C3 MSCs (b).
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