Figure 8.

Relaxin reverses contractile phenotypes associated with miR-144-3p. A, transfection of donor lung fibroblasts with miR-144-3p mimic to decrease RXFP1 levels followed by incubation with low and high concentrations of relaxin. Right panel, transfection of IPF lung fibroblasts with anti-miR-144-3p followed by incubation with low and high concentrations of relaxin. Cells were lysed for immunoblotting for RXFP1, phospho-MLC20, total MLC20, α-SMA, and β-Actin (n = 3). B-E, densitometry of immunoblotting presented in A. Data were expressed as mean ± S.D., normalized to β-Actin or total MLC20. B, between mimic and no mimic, significant effects (p < 0.05) were observed for vehicle, relaxin low, and relaxin high in donor lung fibroblasts. More α-SMA protein was detected in donor lung fibroblasts following transfection of miR-144-3p compared with scrambled control. C, in IPF lung fibroblasts, no significant effects were observed with either low or high dose relaxin between mimic or no mimic groups. D, significant effects between mimic and no mimic were observed only with high dose of relaxin (p < 0.05). Significantly more phospho-MLC20 was present in IPF lung fibroblasts compared with donor lung fibroblasts at baseline. E, significant effects were observed only for relaxin high dose in IPF lung fibroblasts following transfection with antagomir (p < 0.05; n = 3). Data were analyzed by two-way ANOVA followed by the Holm-Sidak test.