Fig. 8

Elevated hepatic oxidative damage in rt269I HBV infection and its clinical implications. A Western blot analysis of p-eIF2α, ATF4, LC3B, and GAPDH in HepG2 cells transfected with mock, rt269L, or rt269L/HBx∆ vector. B ATF4 expression determined by immunofluorescence assay. C, D Measurement of LC3 puncta after cotransfection of the EGFP-LC3 vector with mock, rt269L-, or rt269L/HBx∆-containing plasmids, as determined by immunofluorescence and flow cytometry assays. LC3 is recruited to autophagosomes, forming a punctate structure, as shown by green dots. E Formation of 8-OHdG in hepatocytes transfected with rt269I. HBV representative immunofluorescence images showing 8-OHdG in HepG2 cells transiently transfected with the mock, rt269L, or rt269I vector. Nuclei were stained with DAPI (blue). 8-Hydroxy-2′-deoxyguanosine (8-OHdG). F Detection of 8-OHdG release in serum in mice hydrodynamically infected with mock, rt269L, or rt269I vectors. G Measurement of the 8-OHdG release level in patient serum collected from two independent cohorts (KU and SNU3). H Mitochondrial damage biomarkers in patient serum. Detection of nuclear DNA (nDNA) fragmentation release in serum detected by ELISA and mitochondrial DNA release in serum detected by RT–qPCR. Scatter dot plots were generated using GraphPad Prism 9.0 software (GraphPad, La Jolla, CA, USA). *p < 0.05, **p < 0.01, ***p < 0.001