Figure 3.

miR-144-3p targets RXFP1 in a dose-dependent manner. A, donor lung fibroblasts were transfected with increasing concentrations of miR-144-3p mimic (0.1, 1, 5, and 10 nm). miR-144-3p mimic decreased the expression of RXFP1 and up-regulated the expression of α-SMA in a dose-dependent manner. Densitometry of RXFP1 (B) and α-SMA (C) from A (data analyzed by one-way ANOVA, for RXFP1, p value for trend 0.0006, R² = 0.640) and for α-SMA (p value of log-transformed data for trend <0.02, R² = 0.51). D, top panel, prediction of the seed region of hsa–miR-144-3p WT and mismatch mutant targeting the 3′ UTR of human RXFP1. Bottom panel, representative image of gel contraction experiments of donor lung fibroblasts following infection with WT miR-144-3p, mismatch miR-144-3p, or GFP-expressing particles or incubation with TGFβ (n = 3). E, quantification of collagen gel areas was performed using ImageJ and plotted (p < 0.001, vehicle + miR-144-3p versus vehicle + GFP; p < 0.05, TGFβ + miR-144-3p versus TGFβ + GFP; ns, untreated GFP versus untreated mismatch group). Values are presented as mean ± S.D.