Capza1

Huh7 and HepG2 cells were lysed with RIPA buffer (Beyotime, China) containing protease inhibitors or phosphatase inhibitors (Beyotime, China) for 30 min on ice. The cell lysate was centrifuged at 13000 ×g for 15 min at 4°C, and the supernatant was collected. F-actin (cytoskeletal proteins) were extracted followed by Extraction of cytoplasmic and cytoskeletal proteins. The lysate was heated at 100°C in a metal bath for 5 mins after mixing with 5× loading buffer. Proteins were separated by SDS-PAGE and then transferred to PVDF membranes (Millpore, USA). A solution of 5% skimmed milk was used to block the membranes at room temperature for 1 h and then incubated with a primary antibody at 4°C overnight. The next day, after washing the membranes three times with TBST, the PVDF membranes were incubated with homologous HRP-conjugated secondary antibody at room temperature for 1 h. Finally, the blots were visualized with ECL reagent using an imaging system (Vilber, France). Antibody descriptions were as follows: (FAM21C:1:1000, Millpore, USA; GAPDH:1:10,000, Proteintech, China; HA:1:1000, Roche, Switzerland; F-actin:1:1000, Abcam, USA; CAPZA1:1:5000, Abcam, USA; Flag:1: 1000, Sigma, USA).

A549 cell line, H7N9 and H1N1pdm09 influenza virus were obtained from Shenzhen Center for Disease Control and Prevention (SZCDC). All the reagents for DIGE were purchased from GE Healthcare (Pittsburgh, PA, USA). DL-Dithiothreitol (DTT), Iodoacetamide (IAA), CHAPS were purchased from Sigma–Aldrich (Louis, MO, USA). The antibodies against CAPZA1, OAT, PCBP1, EIF5A, PAFAH1B2 were purchased from Abcam (Cambridge, MA, USA). The antibody against β-action was purchased from Santa Cruz (Santa Cruz, CA, USA). The secondary antibodies of goat anti-mouse and goat anti-rabbit were purchased from Thermo Fisher (Rockford, IL, USA). The restriction enzyme (BamH I and Not I) were purchased from NEB(USA).
Lipofectamine2000 and pcDNA3.1/Neo(+) were purchased from Invitrogen (Carlsbad, CA, USA).