Figure 8.

Dicer1 knockdown in fibroblasts decreases mature miR-29 (microRNA-29) abundance on control extracellular matrix. Fibroblasts were transduced with Dicer1 shRNA or scrambled control to establish stable expression. (A) Shown is an immunoblot for Dicer1. (B and C) Equal numbers of transduced cells were cultured on control extracellular matrix for 18 hours. Medium was removed and equal volume of serum-free medium was added to each reaction for 8 additional hours. (B) Quantitative PCR for mature miR-29a, -29b, and -29c normalized to miR-451. Data are shown as a box-and-whisker plot, and P value was calculated using the Student’s two-tailed t test. (C) Equal volumes of conditioned medium were analyzed by immunoblot for collagen I and MMP-2 (n = 2, representative experiment shown in triplicate). Densitometry values are shown in the lower panel, with error bars representing the SD, and P value was calculated using the Student’s two-tailed t test. *P < 0.01, **P < 0.001, ***P < 0.0001. KD = knockdown.