Anti hbc

Cells were lysed in 1 ​× ​Lysis buffer with proteinase inhibitor at 4 ​°C for 30 ​min. The supernatants of cell lysates were collected, added with loading buffer and incubated at 100 ​°C for 10 ​min. Then proteins of different molecular weight were separated by SDS‒PAGE and transferred onto PVDF membrane (Invitrogen). The membrane was blocked with 5% nonfat milk in TBST (Tris-buffered saline containing 0.1% Tween 20) at room temperature for 1–2 ​h and hybridized with primary antibodies overnight at 4 ​°C. Second antibodies were fluorescent-conjugated or HRP-linked anti-rabbit/mouse IgG. The proteins were finally visualized by Odyssey infrared Imager (LI-COR Biosciences, Nebraska, USA) and Tanon-5200 Chemiluminescent Imaging System (Tanon Science & Technology, Shanghai, China). The primary antibodies were as follows: anti-β-Tubulin (Applygen, Beijing, China), anti-HBc (gifted by Prof. Ningshao Xia of Xiamen University), anti-HBs (Abcam, Cambridge, UK), anti-Flag (Sigma, Missouri, USA). Due to the overlapped amino acid sequences of p22cr with HBc, the antibody of HBc could also detect p22cr via cross-reaction.