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. 2022 Jan 27;13:773001. doi: 10.3389/fimmu.2022.773001

Figure 3.

Figure 3

IFN-γ upregulates XOR expression by stimulating expression and activation of the STAT1. (A-D) HepG2 and Bel-7402 cells were incubated with varying concentrations of IFN-γ for varying time intervals and then XOR mRNA and protein expression were measured by qPCR and western blotting respectively. (E, F) HepG2 and Bel-7402 cells were treated with IFN-γ (10 ng/ml) for 24 h and then XOR activity and intracellular UA content were measured by enzyme coupling assay. (G) The protein expression levels of XOR, IFNGR1, STAT1 and STAT3 were examined by western blotting and (I) the enrichment of STAT1 and STAT3 were assessed by ChIP-qPCR. (H) The effect of IFN-γ (10 ng/ml) on the expression of XOR, IFNGR1 and STAT1 proteins were examined after pre-treatment with cycloheximide (CHX) (500 nM) for 24h. (J, K) After transfection with siRNA for 48h, HepG2 cells were treated with IFN-γ (10 ng/ml) and then the expression of XOR and STAT1 proteins were determined by western blotting. (K) The enrichment of STAT1 at the XDH promoter were assessed by ChIP-qPCR. Significance for all data determined by the independent samples t-test. Data are shown as mean ± S.D., *p < 0.05, **p < 0.01. (E) *p < 0.05 compared with control XOR, # p < 0.05, ## p < 0.01 compared with control XDH. (K) *p < 0.05 compared with control siNC (negative control siRNA); # p < 0.05 compared with IFN-γ siNC.