Fig. 4
UC-MSC-CM inhibits MRTF-A nuclear localization in HEFs. HEFs were treated with TGF-β1 (5 ng/mL) and cultured with or without UC-MSC-CM for 48 h and then stained with MRTF-A antibody (A), rhodamine-phalloidin (C), and counterstained with DAPI. (A) (a–c) no treatment; (d–f) treatment with TGF-β1; (g–i) treatment with TGF-β1 and UC-MSC-CM; (j–l) enlarged images of the region within the white box in (a), (d) and (g), respectively. (B) Nuclear-to-cytoplasmic ratio of MRTF-A was determined for each condition as described in “Methods” section. (D) The fluorescence intensity ratio of F-actin to DAPI was quantified and plotted using ImageJ software: Measurements were taken across 21 distinct same sized surface areas (3900 μm 2 for images taken with zoom factor 1) from 3 independent experiments. Data are expressed as the mean ± SEM. #P < 0.05 and ###P < 0.001 versus the control; **P < 0.01 versus TGF-β1 treatment only (ANOVA w/ Tukey). NS not significant (Mann–Whitney U test); Scale bars, 100 μm; original magnification, ×200.
