Fig. 1: IFIT2 and IFIT3 act together to inhibit viral replication.
a, Human A549 cells were treated with a non-targeting control siRNA (NC, black circles) as a negative control, or siRNA targeting IFIT1 (light grey squares), IFIT2 (blue triangles), IFIT3 (yellow triangles), or IFIT2 and IFIT3 in combination (green diamonds). In indicated conditions, cells were induced with IFNα (1,000 U ml−1). All cells were infected with VSV-GFP (0.01 MOI), and supernatant was collected at 16 hpi for titration (n = 3 biological replicates). p.f.u.s, plaque-forming units. b, Inducible Flp-In T-REx HEK293 cells expressing no IFIT (black circles), mouse IFIT2 (blue squares), mouse IFIT3 (yellow triangles), or mouse IFIT2 and IFIT3 together (green triangles) were mock treated or treated with doxycycline (500 ng μl−1) for 24 h and then infected with VSV-GFP (0.01 MOI). Supernatant was collected at 24 hpi for titration (n = 3 biological replicates). c–e, Using the same cell lines as in Fig. 1b, each cell line was treated with doxycycline (500 ng μl−1) for 24 h and infected with VSV-GFP (3.0 MOI). c, Images were taken at 0, 2, 4, 6 and 8 hpi. Scale bar, 1,000 μm. d, Images from experiments shown in c were processed and quantified for GFP fluorescence intensity (n = 8 biological replicates). e, Lysates from infected cells were collected at 0, 2, 4, 6 and 8 hpi and analysed by western blotting for expression of two viral proteins, VSV-G and VSV-N, and a loading control, GAPDH. All experiments were performed with 3 or more biological replicates with individual datapoints shown (a,b,d) or representative images shown (c). Data are represented as mean ± s.e.m. Statistical analyses: ordinary two-way analysis of variance (ANOVA) with Tukey’s post test and a single pooled variance (a,d); ****P < 0.0001; NS, not significant; ordinary two-way ANOVA with Šídák’s post test and a single pooled variance (b); ****P < 0.0001.
