4 6 diamidino 2 phenylindole dapi staining solution

Hepatocytes were plated on circular coverslips in the wells of the 24-well microplate. After the indicated treatment, the cells were fixed with 4% paraformaldehyde (Sigma-Aldrich) for 15 min at room temperature. After 3 gentle washes with PBS, the cells were incubated with 0.3% Triton X-100 (Solarbio) for 15 min at room temperature to improve permeability. Subsequently, 3% BSA served as the blocking agent to block the hepatocytes for 1 h at room temperature. Then, the coverslips were incubated with the primary antibodies at 4°C for 16 h. The cells were then incubated with the fluoresceine isothiocyanate (FITC)-or Cy3-labeled secondary antibodies for 1 h at 37°C in the dark. After 3 gentle washes with PBS containing Tween 20 (PBST), 4',6-diamidino-2-phenylindole (DAPI) staining solution (Solarbio) was used to counter-stain nuclei in the dark for 10 min. The coverslips were mounted on glass slides using antifade mounting medium and visualized under a laser scanning microscope (LSM) 710 confocal laser microscope system (Zeiss). The primary and second antibody details are listed in the Supplemental Table S2.