Anti skp2

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).

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.