6 diamidino 2 phenylindole dapi

Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL; Roche) assay was carried out to assay the level of hepatocyte apoptosis in liver tissue according to the manufacturer’s instructions. Briefly, after deparaffinized, rehydrated, and retrieved, liver sections were treated with 0.1% Triton X-100 for 8 min at room temperature, blocked with 3% H₂O₂, washed in PBS for 3 times, and then placed in a working solution (10% enzyme solution and 90% label solution) for 1 h at 37 ℃. 4’, 6-diamidino-2-phenylindole (DAPI; Solarbio, China) was added, and the apoptotic cells were observed under a light microscope.

To clarify the location of AdMx protein in the cell, GSM cells (2 × 10⁵ cells/mL) were cultivated on glass coverslips in 12-well culture plates for 24 h until the cells reached approximately 70%–80% confluency. Then, 250 μL M199 medium containing 2 μg plasmid and 5 μL lipofectamine^TM 2000 (Invitrogen, Carlsbad, CA, USA) were introduced into per well according to the manufacturer’s instructions. The medium was changed to fresh M199 that contained 10% FBS after 6 h. At 48 h post transfection, the cell culture media was removed, and the GSM cells were washed with PBS three times, and then fixed with 4% paraformaldehyde for 20 min. After being washed three times with PBS, the cells were stained with 6-diamidino-2-phenyli-ndole (DAPI) (Solarbio, Beijing, China). Lastly, the coverslips were washed and examined using a fluorescence microscope (Olympus, Tokyo, Japan).

MDPCs fixed with 4% paraformaldehyde were permeabilized with 0.1% Triton X‐100 in PBS for 20 min, rinsed with TBS and blocked with PBS containing 5% bovine serum albumin (BSA, Sigma‐Aldrich) for 1 h. The cells were then incubated with primary antibodies at 4°C overnight, followed by washing with PBS. Subsequently, the cells were stained with fluorophore‐conjugated secondary antibodies for 1 h at room temperature and 4′, 6‐diamidino‐2‐phenylindole (DAPI, Solarbio) for 5 min. The results were visualized using an IX73 inverted microscope (Olympus). Fluorescence intensity was quantified using ImageJ.
All antibodies used for immunofluorescence characterizations in this study are listed in Table 2.