Anti myh11

The following antibodies were used for Western blotting and immunofluorescence analyses: anti-GAPDH (CWBIO, Beijing, China, CW0100 M; WB, 1:1000), anti-Acta2 (HUABIO, Hangzhou, China, ER1003; IF, 1:100; WB, 1:1000), anti-Myh11 (Proteintech, Wuhan, China, 60222-1-lg; IF, 1:100; WB, 1:5000), anti-PCNA (HUABIO, Hangzhou, China, ET1605-38; IF, 1:100; WB, 1:1000), anti-MMP2 (Thermo Fisher Scientific, Shanghai, China, 436000; IF, 1:100; WB, 1:1000), anti-4-HNE (R & D Systems, Shanghai, China, MAB3249; IF, 1:50; WB, 1:1000), anti-Nrf2 (Abcam, Cambridge, UK, ab31163; IF, 1:100; WB, 1:1000), anti-TNF-α (Santa Cruz Biotechnologies Inc., CA, USA, sc-52746; IF, 1:200; WB, 1:1000), anti-IL-1β (Santa Cruz Biotechnologies Inc., CA, USA, sc-515598; IF, 1:200; WB, 1:1000), anti-PI3K (CST, Shanghai, China, #4249; WB, 1:1000), anti-p-PI3K (CST, Shanghai, China, #17366; WB, 1:1000), anti-Akt (CST, Shanghai, China, #4685; WB, 1:1000), anti-p-Akt (CST, Shanghai, China, #4060; WB, 1:1000).

Western blot analysis was carried out as described previously (28 ). Protein extracts from cultured MOVAS-1 cells or mouse carotid artery samples were prepared with Western-IP lysis buffer (Beyotime) supplemented with proteinase inhibitor cocktail (Roche). The primary antibodies used for Western blotting include: anti-AGGF1 (Proteintech, 1189-1, 0.7 μg/ml), anti-MYH11 (Proteintech, 21404-1, 0.7 μg/ml), anti-α-SMA (Proteintech, 55135-1, 0.6 μg/ml), anti-SM22 (Proteintech, 10493-1, 0.33 μg/ml), anti-GAPDH (Proteintech, 60004-1, 0.166 μg/ml), anti-pMEK1/2 (Bioss, bsm-52176R, 1 μg/ml), anti-pELK (Bioss, bs-10154R, 1 μg/ml), anti-ERK1/2 (Abcam, 184699, 1.2 μg/ml), anti-pERK1/2 (Abcam, 223500, 1:1000 dilution), anti-integrin α5 (Proteintech, 10569-1, 0.5 μg/ml), anti-integrin α7 (Abclonal, A14246, 1:500), and anti-integrin α8 (Abclonal, A13056, 1:500). The secondary antibodies include a goat anti-rabbit IgG or a goat anti-mouse IgG (HRP-conjugated, 1:20,000) from Biofly. Images from Western blot analysis were captured using a ChemiDoc XRS (Bio-Rad Laboratories) with the SuperSignal West Pico Chemiluminescent Substrate (Pierce Chemical Co) and further analyzed with Gel-Pro analyzer.