Af7021

Western blots were performed as previously described [39 (link)]. The primary antibodies were FADS1 (1:1000; ab126706), FADS2 (1:1000; Proteintech 28034-1-AP), GAPDH (1:5000; Affinity AF7021), actin (1:5000; Proteintech 60008-1-Ig). Then, the corresponding secondary antibodies were incubated at room temperature for 1 h. The bound antibody was expressed using the ECL method and detected with the BIO-RID imaging system (BIO-RAD, Hercules, CA, USA). The grayscale values of all bands were analyzed with Image Lab (Image J 1.53C, Millipore, Burlington, MA, USA) software.

Total protein was extracted from human NP cells by RIPA lysis buffer containing protease inhibitor (Beyotime). Nuclear and cytoplasmic proteins were extracted from NP cells by a Nuclear‐Cytosol Extraction Kit (Solarbio) according to the manufacturer's instructions. Western blot procedure was performed as previously described.³³ Primary antibodies against the following proteins were used: myosin IIA (Abcam, ab138498, 1:1000), myosin IIB (Abcam, ab230823, 1:1000), p21 (CST, 2947, 1:1000), p53 (Proteintech, 10442‐1‐AP, 1:1000), Cyclin D1 (Abcam, ab134175, 1:10 000), CDK4 (Abcam, ab108357, 1:1000), MRTF‐A (Abcam, ab115319, 1:1000), RhoA (Abcam, ab187027,1:1000), p‐RhoA‐Ser188 (Abcam, ab41435, 1:1000), ROCK1 (Proteintech, 21850‐1‐AP, 1:1000), ROCK2 (Proteintech, 20248‐1‐AP, 1:1000), MLC (CST, 3671, 1:1000), p‐MLC‐Ser19 (CST, 3671, 1:1000), GAPDH (Affinity, AF7021, 1:2000), Lamin B (Proteintech, 12987‐1‐AP, 1:1000). GAPDH and Lamin B were used for normalization. The experiments were repeated three times.