Abcc10

Total cell lysates were collected and protein concentration was measured. Equal amount of proteins was separated by SDS-PAGE and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA). The membranes were blocked with 5% bovine serum albumin in TBST for 2h at room temperature and incubated with primary antibodies overnight at 4°C. Following the incubation with secondary antibodies at room temperature for 2h, proteins on the membrane were visualized with a chemiluminescence kit (Thermo Scientific). Primary antibodies are listed as follows: β-actin (1:1000, Cell Signaling Technology), FOXM1 (1:100, Santa Cruz Biotechnology), cleaved caspase-3 (1:1000, Cell Signaling Technology), cleaved caspase-7 (1:1000, Cell Signaling Technology), cleaved PARP (1:1000, Cell Signaling Technology) and ABCC10 (1:50, Santa Cruz Biotechnology).

Tissue slides were deparaffinized, rehydrated, followed by antigen retrieval. After the incubation of primary and secondary antibody, the slides were incubated with diaminobenzidine (DAB) (Dako, USA), and finally counterstained with hematoxylin (Sigma Chemical Co, USA). Primary antibodies are listed as follows: Ki67 (1:500, Abcam), FOXM1 (1:100, Santa Cruz Biotechnology), ABCC10 (1:25, Santa Cruz Biotechnology)