FIG. 2. miR-29s downregulated MMP2 and reduced MMP2 activity. (A) HT1080 cells were transfected with control miRNA or miR-29s mimics (50 nM) for 72 h. For western blotting, cells lysates were prepared and analyzed using the indicated antibodies. Actin was used as a loading control. (B) After 72 h, conditioned medium from the same experiment was harvested and used for western blotting. (C) Conditioned medium from the same experiment was processed for gelatin zymography. The band at 72 kDa represents the active form of MMP2, and the band at 92 kDa represents pro-MMP2. (D) Quantitative real-time PCR. HT1080 cells were transfected with 50 nM of miR-29s mimics. After incubation for 48 h, total RNA was isolated and analyzed by SYBR quantitative real-time PCR. The results were normalized to 18s RNA expression and are presented as the relative expression level (*p<0.05).
