Orai1

The cells were lysed by lysis buffer (Boster, Wuhan, China) containing 1 mM protease inhibitor PMSF (Boster, Wuhan, China). To obtain total protein, the lysate was centrifuged at 13,000 rpm for 15 min at 4°C. The supernatant was the total protein. The total protein was denatured and then separated by SDS-PAGE (8–15%). The separated protein was transferred to a nitrocellulose membrane (Univ-bio, Shanghai, China). The membrane was blocked with a protein free quick blocking solution (Beyotime, Shanghai, China) for 15 min and incubated with specific primary antibodies for 3 h at room temperature. The membrane was washed for 3 times with Tris-buffered saline with 1% Tween 20 (TBST) (Beyotime, Shanghai, China) and incubated with secondary goat anti-rabbit IgG (1:5000, BA1056, Boster, Wuhan, China) for 1 h at room temperature. The membrane was washed again for 3 times with TBST, and the blot was detected by enhanced chemiluminescence solution (Meilune, Dalian, China) and analyzed by Image J (version 1.4.3.67) software. The primary antibodies used are as follows: β-actin (1:5000, bsm-33036M, Bioss, Beijing, China), GRP78 (1:1000, bs-1219R, Bioss, Beijing, China), CHOP (1:500, bs-20669R, Bioss, Beijing, China), STIM1 (1:500, A9764, Abclonal, Wuhan, China), ORAI1 (1:500, A7412, Abclonal, Wuhan, China).