Sab1412855

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).

Cell Lines, Antibodies, and Other Reagents Seven human ESCC cell lines (TE2, TE4, TE5, TE8, TE9, TE13, and TE15) and the PK59 human pancreatic cell line were obtained from the Riken Cell Bank (Tsukuba, Japan), and four other human ESCC cell lines (KYSE70, KYSE150, KYSE170, and KYSE790) were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). Two cell lines (TE15, KYSE170) were used for in vitro studies, with these cells being cultured in RPMI-1640 medium (Nacalai Tesque, Kyoto, Japan) supplemented with 100 U/mL penicillin, 100 mg/mL streptomycin, and 10% fetal bovine serum under a humidified 5% CO 2 atmosphere at 37 C. 10e18 For immunohistochemistry, a polyclonal anti-LRRC8A antibody (ab157489) and a monoclonal anti-cytokeratin 13 antibody (ab16112) were purchased from Abcam (Cambridge, MA), whereas the monoclonal anti-LRRC8A antibody (SAB1412855) for the protein assay was from Sigma-Aldrich (St. Louis, MO), and the mouse monoclonal antibody that targeted b-actin was also from Sigma-Aldrich. Horseradish peroxidaseeconjugated anti-rabbit or mouse secondary antibodies were from Cell Signaling Technology (Beverly, MA).