Monoclonal anti gsk 3β antibody

The human ESCC cell lines TE2 and TE5 were obtained from the Cell Resource Center for Biomedical Research at the Institute of Development, Aging, and Cancer (Tohoku University, Sendai, Japan). These cells were grown in RPMI-1640 medium (Nacalai Tesque, Kyoto, Japan) supplemented with 100 U/mL of penicillin, 100 μg/mL of streptomycin, and 10% fetal bovine serum (FBS). Cells were cultured in flasks and dishes in a humidified incubator at 37°C under 5% CO₂ in air. The polyclonal anti-NHE1 antibody used for the immunohistochemical analysis and protein assay was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The following antibodies were used in the Western blot analysis; a monoclonal anti-AKT antibody, monoclonal anti-phospho AKT antibody, monoclonal anti-GSK-3β antibody, monoclonal anti-Phospho-GSK-3β antibody, polyclonal anti-β-catenin antibody, and monoclonal anti-p21 antibody were from Cell Signaling Technology, and a monoclonal anti-β-actin antibody and a monoclonal anti-p53 antibody were from Sigma-Aldrich (St. Louis, MO, USA). The following antibodies were used in the immunohistochemical staining; polyclonal anti-β-catenin antibody was from Cell Signaling Technology, and anti-Snail goat polyclonal antibody was from Abcam.

Fruitflow was provided by By-Health Co., Ltd. (Zhuhai, Guangdong, China). Aspirin (acetylsalicylic acid) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Collagen and ADP were purchased from Chrono-Log (Havertown, PA, USA). Human platelet factor 4 (PF4; CXCL4), thromboxane B₂ (TXB₂), and 6-keto-prostaglandin F_1α (PGF_1α) enzyme-linked immunosorbent assay (ELISA) kits were purchased from Abcam (Boston, MA, USA). Polyclonal anti-Akt antibody and monoclonal anti-Syk antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Monoclonal anti-phospho-Akt (Ser473) antibody, monoclonal anti-GSK3β antibody, monoclonal anti-phospho-GSK3β (Ser9) antibody, polyclonal anti-p38 mitogen-activated protein kinase (MAPK) antibody, polyclonal anti-phospho-p38 MAPK (Thr180/Tyr182) antibody, monoclonal anti-phospho-PLCγ2 (Tyr759) antibody, and polyclonal anti- PLCγ2 antibody were purchased from Cell Signaling Technology (Danvers, MA, USA). Monoclonal anti-phospho-syk (Tyr525) were purchased from Abcam (Boston, MA, USA). Phalloidin-iFluor 555 was purchased from Abcam (Boston, MA, USA), which is one of a series of phalloidin conjugates that bind to actin filaments.