Ab74793

Proteins were extracted from cells or tissues and added to the RIPA buffer. Cracking on ice for 30_60 minutes. The supernatant was collected by centrifugation at 4°C and 12000 rpm for 15 minutes. BCA protein assay kit (Boster Inc, China) was used for protein quantification. SDS-PAGE gel electrophoresis was performed. Then, the protein was transferred to the nitrocellulose membrane and 5% skimmed milk powder blocked the nonspecific antigen. The primary antibody was added and incubated at 4°C overnight. After washing the membrane, horseradish peroxidase-labeled IgG (secondary antibody) was added at 37 °C for 1h. Densitometric analysis was performed by Image Lab. In the assay, antibodies included rabbit anti-hnRNP E1 (Abcam, UK. ab74793, 1:1000), mouse anti-HPV16 E2 (Abcam, UK. ab17185, 1:1000), mouse anti-HPV16 E6 (Abcam, UK. Ab70, 1:1000), mouse anti-β-actin (Boster Inc, China, 1:300).

Proteins were extracted from cells and animal tissues. Next, the proteins were transferred onto polyvinylidene fluoride membranes (Millipore, Bedford, MA) using electrophoresis and transferred to a 0.2-µm nitrocellulose membrane (GE Healthcare, Chicago, IL USA). Membranes were blocked with 5% nonfat milk and incubated with primary antibodies overnight at 4 ℃. The primary antibodies used for western blotting were as follows: PCBP1 (1:2000, ab74793, Abcam), DKK1 (1:1000, ab109416, Abcam), phosphor-β-catenin (1:1000, 9561S, CST), Claudin-1 (1:1000, 13255 s, CST), β-catenin (1:1000, 8480 s, CST), and Vimentin (1:1000, 5741 s, CST). The signalling detection was performed using an ECL detection kit (WBKLS0500, Millipore).