Fig. 2

The cytotoxic in vitro effect of BVDV on HMCLs. a Mean ± SD of the percentage of Apo 2.7⁺ cells in JJN3, MM1.S, OPM2, and NCI-H929 after 24, 48, and 72 h of treatment with BVDV (1 MOI). The graphs represent the mean percentage of Apo2.7⁺ cells of four independent experiments for each cell line evaluated by flow cytometry. b Representative histogram plots of flow cytometry showing the percentages of NCI-H929 cells positive for the apoptotic marker APO2.7, after 24, 48, and 72 h of BVDV (1 MOI) treatment or in the control condition. c Pro- and active-caspase 3 expression was evaluated by western blot in JJN3 and OPM2 cells treated with or without BVDV (1 MOI) for 48 h. β-actin was used as loading control and JJN3 treated with high doses of Bor as positive control (Cnt+). The histogram represents the protein bands intensity quantified using ImageJ software reported as arbitrary unit normalized by the loading control. d Western blot of Bcl-2 and Mcl-1 expression on JJN3 and OPM2 cells treated for 48 hours with or without BVDV (1 MOI). β-actin was used for loading control and RPMI-8226 cells line as positive control for both protein (Cnt+). The histograms represent the protein bands intensity quantified using ImageJ software reported as arbitrary unit normalized by the loading control. The p values were calculated by two-tailed Student’s t test. (*p < 0.05, **p < 0.01, ***p < 0.001) (CNT = control, untreated cells)