Journal: Chinese Medical Journal
Article Title: Follistatin-Like 1 Promotes Bleomycin-Induced Pulmonary Fibrosis through the Transforming Growth Factor Beta 1/Mitogen-Activated Protein Kinase Signaling Pathway
Figure Lengend Snippet: FSTL1 modulates myofibroblast differentiation by facilitating p38/JNK signaling. (a and b) Primary lung fibroblasts from Fstl1+/− and their WT littermates were treated with 5 ng/ml TGF-β1. (a) Immunofluorescence staining of α-SMA in lung fibroblasts. (b) Protein expression levels of α-SMA and type I collagen. (c-e) MLgs were pretreated with U0126, SB202190, and SP600125. Protein expression levels of α-SMA and type I collagen were detected by Western blotting. n = 4; Bars = 100 μm. * P
Article Snippet: MLgs were cultured in 96-well plates for 24 h, and serum starved for 24 h. The cells were preincubated for 1 h with U0126 (ERK inhibitor, Cell Signaling Technology, MA, USA), SB202190 (p38 inhibitor, Cell Signaling Technology, MA, USA), SP600125 (JNK inhibitor, Cell Signaling Technology, MA, USA), and SB525334 (Smad2/3 inhibitor, R and D Systems, MN, USA) before treatment with 5 ng/ml TGF-β1 (R and D Systems, MN, USA) in the presence or absence of 100 ng/ml recombinant human FSTL1 protein (R and D Systems, MN, USA) for 24 h and finally incubated with MTT (final concentration of 0.5 mg/ml) for 4 h. The supernatant was removed, and 150 ml/well of dimethylsulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) was added to dissolve the blue formazan crystals by shaking the plates for 15 min on an orbital shaker at 25°C.
Techniques: Immunofluorescence, Staining, Expressing, Western Blot