Anti phospho p53 thr81

The human GC cell lines MKN7, MKN45, MKN74, HGC27, and NUGC4 were obtained from the Riken Cell Bank (Tsukuba, Japan). These cell lines were cultured in RPMI-1640 (Nacalai Tesque, Kyoto, Japan) supplemented with 100 µg/ml of streptomycin, 100 U/ml of penicillin (Nacalai Tesque), and 10% fetal bovine serum (FBS) (Sigma-Aldrich Japan, Tokyo, Japan) in a humidified incubator at 37 °C in 5% CO 2 . The rabbit polyclonal anti-LRRC8A antibody used in the immunohistochemical (IHC) analysis was purchased from Abcam (ab-157489, Cambridge, MA, UK). The mouse monoclonal anti-LRRC8A antibody used in western blotting was purchased from Sigma-Aldrich (SAB1412855 St. Louis, MO, USA). A mouse monoclonal anti-beta-actin (ACTB) antibody was purchased from Sigma-Aldrich (A5441). Rabbit polyclonal anti-SAPK/JNK, anti-phospho-SAPK/JUN, and anti-phospho-p53 (Thr81) antibodies, a mouse monoclonal anti-p53 (1C12) antibody, and a rabbit anti-p21 (Waf1/ Cip1) antibody were purchased from Cell Signaling Technology (anti-SAPK/JNK; #9252/anti-phospho-SAPK/JNK; #9251/anti-phospho-p53 (Thr81); #2676/anti-p53 (1C12); #2524/anti-p21 (Waf1/Cip1); #2947, Beverly, MA, USA). Anti-horseradish peroxidase (HRP)-linked anti-mouse and anti-rabbit secondary antibodies were obtained from Cell Signaling Technology (anti-mouse; 7076S/anti-rabbit; 7074S).