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. 2021 Jun 29;11:13484. doi: 10.1038/s41598-021-92871-z

Figure 6.

Figure 6

TGF-β1 concurrently activated SMAD2/3 and YAP/TAZ in HVOX fibroblasts. HVOX fibroblasts were treated with 10 ng/mL TGF-β1 for 0, 15, and 30 min and 1, 2, 3, 4, 6, 12, and 24 h (a,b). Distributional changes of YAP, TAZ, and SMAD2/3 were assessed by immunostaining (a; bar = 50 µm). Western blotting was performed to detect Ser465/467-phosphorylated SMAD2 (pSMAD2), SMAD2, Ser423/425-phosphorylated SMAD3 (pSMAD3), SMAD3, Ser127-phosphorylated YAP (pYAP), YAP, Ser89-phosphorylated TAZ (pTAZ), TAZ, and β-actin in the total cell fraction (b). HVOX fibroblast were treated with 10 ng/mL TGF-β1 for 0, 15, and 30 min and 1, 3, 6, and 24 h (c). Western blotting was performed to detect SMAD2, SMAD3, YAP, TAZ, GAPDH, and lamin A/C in the cytoplasmic (C) and nuclear (N) fractions. In (b) and (c), representative grouping of gels cropped from different parts of the same gel or from different gels; whole gels available in supplemental material.