Fig 2: QA treatment inhibits ROS production by HSV-1 infection in SK-N-SH cells. (A) SK-N-SH cells were infected with HSV-1 (MOI = 0.01) for 2 h and then treated with QA at 50 and 100 µg/mL and cultured for 30 h. ROS was labeled with DCFH-DA. The level of ROS was measured using flow cytometry. (B-D) SK-N-SH cells were infected with HSV-1 (MOI = 0.01) for 2 h and then treated with QA at 50 and 100 µg/mL and cultured for 48 h. (B) Whole cell extracts were subjected to western blot analysis for TNF-a and IL-6. ß-actin was used as an internal control. (C) RNA was extracted from SK-N-SH cells, and TNF-a and IL-6 expression was measured by real-time PCR. (D) Whole cell extracts were subjected to western blot analysis for p-TBK1, TBK1, p-IRF3, IRF3, and VP16. ß-Actin was used to confirm equal sample loading. The data are representative of three independent experiments and quantified as mean values ± SEM. One-way ANOVA with Tukey’s post hoc test; *** p < 0.001, compared with the HSV-1 treatment.

Fig 3: IRF3 immunostaining of the bile ducts is associated with NRF2 activity and with SOCS3 protein expressions. (A) We depicted positive significant correlations of moderate degree to pNRF2 immunopositivity of the nuclei (r = 0.364; p = 0.006) and (B) SOCS3 cytoplasmic staining (r = 0.368; p = 0.006). Pearson’s correlation coefficient was performed with significance set at p = 0.05.

Fig 4: Associations between IRF3 immunostainings and disease progression. Study of the bile ducts in the cohort depicted that IRF3 staining intensity was in NASH higher than in NAFL (A–D). In NASH, there are more bile ducts having a strong IRF3 staining intensity of +3 than in NAFL; * p = 0.027 (A). The average IRF3 staining intensity of the bile ducts per case was in NASH higher than in NAFL; p = 0.002 (C). Representative IHC images of NAFL (B) and NASH (D) show representative areas of IRF3 immunopositive stained bile ducts (arrows) in NAFL (B) and NASH (D) with stronger IRF3 staining in NASH. Differences between groups were analyzed using Mann–Whitney U tests; bold lines inside the box plot represent median levels. Results are significant at * p = 0.05 and ** p = 0.01; bars = 200 µm.