Fig 1: Cytotoxicity of T1012G against gastric cancer cells, virus replication and expression of IFN-I response genes after T1012G infection with altered STAT3 gene expressions. A and B Cell viability (measured by cck8) of HGC-27 gastric cancer cells transfected with siRNA or plasmid was assayed 2 days after infection of T1012G at different MOI. C After 48 h of incubation with or without propranolol (40 µmol/l), cells transfected with control treatment, siRNA or plasmid were infected with different dose of virus (0.01, 0.05, 0.1, 0.5, 1 MOI). The number of surviving cells in each well was determined 2 days after infection. D, F and H Western blot analysis of STAT3 in the transfected siRNA (si-1 and si-2) and plasmid. E, G and I Cells were infected (MOI 0.01) with HSV-1 (T1012G) and virus yields were determined on Vero cells after 48-h infection. J, L and N Expression of IFN-I responsive genes, PKR, 9 or 20 h following T1012G infection (0.01 MOI) in J and L: HGC-27 cells treating with STAT3 siRNA or vehicle and N: HGC-27 cells treating with plasmid-STAT3 or vehicle. K, M and O Quantification of J, L, N. Results are presented as mean ± SEM, significant differences were evaluated using one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.01 and ****P < 0.0001 (Tukey test for multiple comparisons). **P < 0.01 and ***P < 0.01 (Dunnett's test for multiple comparisons). OE overexpression
Fig 2: Propranolol treatment inhibited the IFN-I-mediated induction of antiviral genes and counteracted IFN-mediated inhibition of viral propagation in HGC-27 human gastric cancer cells. A and B Dose effects of propranolol were evaluated on the IFN-induced inhibition of T1012G production in HGC-27 cells. Cells were treated with 300 ng/well of human recombinant IFN-a/ß and different concentrations (0, 10, 20, or 40 µmol/l) of propranolol for 48 h and infected withT1012G (0.01 MOI). Two days after infection, the cells and medium were harvested, and yields of virus were determined were determined on Vero cells. C and E HGC-27 cells were incubated with 40 µmol/l propranolol (for 48 h) and/or 300 ng/well human recombinant IFN-a/ß (for 6 h) and then harvested for the assessment of protein expression of STAT3, p-STAT3, PKR, and p-PKR by western blot analysis. D and F Quantification of C, E. Results are presented as mean ± SEM, significant differences were evaluated using one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001(Dunnett’s test for multiple comparisons)
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