Anti lamin a c

For immunostaining, cells were washed with PBS three times and then fixed in 4% paraformaldehyde. Cells were frozen in liquid nitrogen and fixed on slides. Slides were blocked in PBSTA [PBS with 0.1% Triton X-100 and 3% bovine albumin (BSA)] for 1 h at room temperature; with primary antibodies, overnight at 4°C; and with secondary antibodies, for 1 h at room temperature. Cell nuclei were stained by using Vectashield mounting medium with 4′,6-diamidino-2-phenylindole (DAPI; H-1800–2, Vector Laboratories). Anti-lamin A/C [rabbit monoclonal antibody (mAb), A19524] was purchased from ABclonal Technology (WuHan, China). Anti-GFP (mouse mAb, ab1218) and anti-fibrillarin (mouse mAb, ab4566) were purchased from Abcam Technology. Immunofluorescence images were collected by using a Zeiss LSM 980 Upright laser scanning confocal microscope with a ×63 objective. Live-cell images were collected by using a Nikon ECLIPSE Ti-E inverted microscope with either ×40 or ×20 objectives.

Total cell lysates were prepared from cultured cells using SDS lysis buffer (Beyotime, China). Mouse liver tissues were lysed in TBS (10 mM Tris-HCl at pH 7.0, 150 mM NaCl) containing 0.5% NP-40 followed by centrifugation. HBV capsids were first separated by electrophoresis in 1% agarose gel and then blotted onto nitrocellulose for western blot analysis as previously described (27 (link)). Nuclear and cytoplasmic fractions were prepared using Nuclear and Cytoplasmic Protein Extraction Kit (Beyotime, China).
Western blot was performed using the following antibodies: anti-Lamin A/C (1:3000, A0249), anti-ANXA1(1:3000, A0249), anti-mouse HMGA1 (1:3000, A4343) from Abclonal; anti-human HMGA1 (1:3000, #7777) and anti-FOXO3α (1:3000, #12829) from Cell Signaling Technology; anti-Flag (1:10000) and anti-β-actin (1:10000) from Sigma; anti-PGC1α (1:3000, 66369–1-lg) and anti-SP1 (1:3000, 21962–1-AP) from Proteintech; anti-HBcAg from DAKO (1:10 000, Copenhagen, Denmark). Mouse anti-HBx mAb 2A7 (1:1000) has been previously reported (28 (link),29 (link)).