Figure 2.

NM-IIA blocking partially reduced the TGF-β-mediated SMA expression and SMA stress fiber formation in fibroblasts. (A) Western blot examination of SMA in NFs and KFs. Fibroblasts were pretreated with 50 μM of blebbistatin (an inhibitor for NM-II) for 24 h followed by cotreatment with 20 ng/mL of TGF-β for 48 h. The GAPDH levels were used as loading controls. (B) is the quantification of Western blotting from (A). (C–F) KFs and NFs were treated with blebbistatin (C, D) or transfected with si-NM-IIA (E, F) for 24 h followed by treatment with TGF-β (20 ng/mL) for 48 h. The fibroblasts were stained with anti-SMA antibody using immunofluorescence examination. White rectangle in each image indicates the enlarged area for high magnification in right-down panel, respectively. The percentage of fibroblasts expressing SMA-mediated stress fibers (C, E) was quantified in (D, F), respectively. The experiments were repeated for at least three times to reach consistency and each experiment was done in triplicates. Color images are available online.