Anti p27

Ham’s F-12K medium, McCoy’s 5A medium, Dulbecco’s modified Eagle’s medium (DMEM), Minimum Essential Medium (MEM), 100 U/mL streptomycin, and 100 μg/mL penicillin were acquired from Gibco (Life Technologies, Waltham, MA, USA). Fetal bovine serum was obtained from BI (Biological Industries, Beit Haemek, Israel). An SRB assay kit was purchased from Bestbio (Shanghai, China). Cell cycle and apoptosis analysis kits were purchased from Yeasen Biotech (Shanghai, China). A Reactive Oxygen Species Assay Kit, cycloheximide (CHX), and Hoechst 33258 were obtained from Beyotime (Beyotime Biotechnology, Shanghai, China). Normal goat serum was purchased from Boster (Boster Biological Technology, Wuhan, China). Anti-Cyclin D1, anti-Cyclin E1, anti-P27, anti-GAPDH, anti-β-actin, anti-FOXO3A, anti-SKP2, anti-γH2AX, HRP-conjugated affinipure goat anti-rabbit, HRP-conjugated affinipure goat anti-mouse, and CoraLite488-conjugated affinipure goat anti-rabbit were purchased from Protein Tech (Proteintech group, Wuhan, China). Ultrapure water was produced with an ultrapure water system (Yipu Yida Technology, Nanjing, China). All of the analytically pure chemicals and solvents were purchased from Sinopharm Group (Shanghai, China).

The following antibodies were used in this study: anti-KAT6A [1:1000 for Western blot (WB), 1:50 for immunohistochemical (IHC) staining, 1:100 for immunoprecipitation (IP), Abnova]; anti-Ki-67 (1:500 for IHC, 1:2000 for flow cytometry, Invitrogen); PCNA (1:5000 for WB, Abcam), anti-Flag (1:1500 for WB, 1:100 for IP, Sigma-Aldrich); anti-HA (1:1000 for WB, 1:100 for IP, CST); anti-Ub, anti-Histone H3 (1:1000 for WB, CST); anti-STAT3, acetyl-Histone H3 (Lys23), acetyl-Histone H3 (Lys14), and acetyl-Histone H3 (Lys9), anti-acetylated lysine (1:1000 for WB, CST); anti-COP1 (1:1000 for WB, 1:200 for IF, 1:100 for IP, Abcam); anti-β-catenin (1:5000 for WB, 1:250 for IF, Abcam); anti-c-Myc, anti-CyclinD1, anti-p27 (1:1000 for WB, Proteintech), anti-p53 (1:5000 for WB, Abcam) anti-ETS2 (1:3000 for WB, Abcam), anti-c-JUN (1:2000 for WB, Abcam), anti-cleaved caspase-3, c-PARP (1:1000 for WB, CST), anti-Puma (1:3000 for WB, Abcam), anti-Bcl2 (1:1000 for WB, Abcam), anti-Bcl-XL (1:1000 for WB, Abcam) and anti-β-actin, anti-GAPDH, (1:1000 for WB, Proteintech). Cisplatin and CHX were obtained from Med Chem Express. WM-1119 and MG132 were obtained from Selleck. Wnt3a was obtained from R&D systems.

All human GC cell lines (BGC823, HGC27, MGC803, MKN45, and SGC7901), normal gastric cell line (GES-1) and human embryonic renal cell line 293FT were obtained from the American Type Culture Collection (ATCC, Beijing, China). All cell lines were tested mycoplasma-negative. MG132 and CHX were obtained from Sigma (Shanghai, China). Anti-ARIH2, anti-p21, anti-p27, anti-CDK1, anti-CDK2, anti-α-Tubulin, anti-HA, anti-SKP2, anti-RNF126 and anti-UHRF2 antibodies were purchased from Proteintech (Wuhan, China). Anti-MYC, anti-Flag, anti-phospho-ATR, anti-phospho-ATM, anti-ɣ-H2AX and anti-cleaved-caspase-3 antibodies were obtained from Cell Signaling Technology (Shanghai, China). Anti-Ki67 and anti-Bcl2 antibodies were purchased from BD Biosciences. All antibodies were diluted according to the manufacturer’s instructions.