Anti zyxin

The following primary antibodies were used (dilution is indicated for immunofluorescence unless stated otherwise): anti-α-actinin (ab18061, Abcam, 1:100 dilution), anti-HA (3F10, Sigma-Aldrich, 1:200 dilution), anti-vinculin (#V9131, Sigma-Aldrich, 1:400 dilution), anti-zyxin (sc-6437, Santa Cruz, 1:40 dilution), anti-talin (#T3287, Sigma-Aldrich, 1:100 dilution), anti-β-actin (#MCA5775GA, Bio-Rad Laboratories, 1:200 dilution, 1:2000 for western blot), anti-γ-actin (#MCA5776GA, Bio-Rad Laboratories, 1:200 dilution, 1:2000 for western blot), anti-actin (#A2066, Sigma-Aldrich, 1:5000 dilution for western blot), anti-myosin IIA (#909802, BioLegend, 1:100 dilution), and anti-phospho-myosin light chain (#3671, Cell Signalling Technology, 1:100 dilution). Secondary antibodies conjugated to Alexa647, Alexa555, or Alexa568 were used (Life Technologies, 1:400 dilution). Actin was stained with Alexa488/Alexa633-conjugated phalloidin (Life Technologies, 1:200 dilution). Blebbistatin was purchased from Sigma-Aldrich (#B0560, 50 μM for 60 min).

The Sects. (4 μm thickness) after deparaffinization were stained with hematoxylin and eosin to observe structural characteristics of renal tissues. Masson and Sirus-red were performed to reflect the deposition of collagens. For immunohistochemical staining, the deparaffinized sections were put in microwave oven to repair antigen. Slides were permeated in 0.1% tritonx-100 and then incubated with anti-LATS2 (Novus, USA), anti-YAP (CST, USA), anti-αSMA (Protein-tech, USA), anti-SIAH2 (Novus, USA), anti-Zyxin (Santacruz, USA) and anti-CTGF (Santacruz, USA) at 4℃ overnight and cover with endogenous peroxidase blockers before incubating with corresponding HRP-conjugated secondary antibodies (Fude-biological, China). Slides were stained with 3,3′-diaminobenzidine (DAB) and then imaged under microscope (Zeiss, Germany).