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. 2016 Mar 1;6:22487. doi: 10.1038/srep22487

Figure 2. HCV infection activated each of the three arms of the UPR in a time dependent manner.

Figure 2

Huh 7.5.1 cells were infected with HCV-JFH1. Following 24, 72, 129, and 168 hours of infection, protein was extracted at the indicated time points. The expression of phospho-IRE1, phospho-PERK, and cleaved ATF6 were analyzed by western blot. JFH1 HCV infection increased levels of phospho-IRE1, phospho-PERK, and the cleaved active form of ATF6 (Fig. 2A). Gene expression levels of spliced XBP-1, ATF3, and ATF4 at day 1, 3, 5, and 7 of infection were determined by real-time PCR and normalized to GAPDH. We found that the spliced XBP-1 mRNA level in JFH1-infected Huh7.5.1 cells increased by 2.7-fold at day 3 post infection compared to uninfected Huh7.5.1 cells (Fig. 2B). The spliced ATF3, and ATF4 mRNA levels were elevated by 20.5, 5.8-fold, respectively, in JFH1-infected Huh7.5.1 cells at day 3 post-infection compared to uninfected Huh7.5.1 cells (Fig. 2C,D). *p < 0.05; **p < 0.01; and #p < 0.001 compared between JFH1-infected and un-infected Huh7.5.1 cells. Empty bar: Huh 7.5.1 cells, Black bar: JFH1-infected Huh 7.5.1 cells.