Figure 2

USP18 deficiency inhibited DENV replication in BMDCs from ifnar1-KO and stat1-KO mice. BMDCs were prepared from ifnar1-KO mice as described in the Materials and Methods. After introduction of siCtrl or siUSP18 via electroporation as described in the Materials and Methods, the cells (1 × 10⁶/ml) were infected with mock or DENV (MOI = 1) for 24 h and then collected for further analysis. The expression of USP18 and NS2B, and protein ISGylation in total cell lysates was determined by Western blotting (A). The levels of DENV RNA were measured by qPCR (B). In addition, the DENV titers were evaluated by plaque assays as described in the Materials and Methods (C). Similar experiments were carried out in BMDCs from stat1-KO mice and the results were presented in (D, E and F), respectively. Values represent the mean of the individual measurements in each sample ± SEM. Statistical analysis was done using Student’s t test (A–F) to compare differences among different treatments. *P < 0.05, **P < 0.01. Original gels are presented in Supplementary Fig. 10.