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. 2016 May 27;6:26827. doi: 10.1038/srep26827

Figure 4. BX795 blocks the phosphorylation and activation of IRF3 leading to suppression of interferon stimulated genes (ISGs).

Figure 4

HepG2/C3A cells, untreated or treated with BX795, were transfected with (a) HEV Rluc RNA, (b) HEV Rluc GAA and (c) double stranded RNA analogue (poly I:C) and processed for immunoblotting for the detection of phosphorylated IRF3 (P-IRF3, Ser396). The blot was reprobed for the detection of total IRF3 (T-IRF3) and actin at indicated time points.The relative protein band density for p-IRF3 was normalized with actin and compared with mock treated and BX795 treated cells at different time points and mentioned as relative fold (RF) values. Cropped blots are used in the main figure and full length blots are included in Supplementary Figure 1. (d–f) HepG2/C3A cells left untreated or treated with BX795 were transfected with HEV Rluc RNA and processed for qRT-PCR of type I IFN genes (IFNA and IFNB) and ISGs (PKR and Mx1) at 24 h post transfection. The data represents mean ± SD of three independent experiments [*, ** and ***represent p < 0.05, p < 0.01 and p < 0.001 respectively, statistical comparisons were done by one-way analysis of variance (ANOVA) between HEV Rluc and BX795 + HEV Rluc].