MDA5 effects on IFN response during HRV infection. (A) “Response to type I interferon” genes (GO term GO0034340) versus all genes. (B) Heatmap representation of RNA-seq data showing HRV-induced IFN-regulated gene transcripts (RPKM) over the course of infection with HRV-B14 (MOI: 1). Data in A show box-and-whisker plots from triplicate infections. A549 cells were previously transfected with MDA5 or nonspecific negative control siRNA (siNeg) and correspond to Fig. 4 F. Mean of triplicate expression ratios from triplicate infections were shown for each time point in B. *, P < 0.05, by Kolmogorov-Smirnov test in A and B; other comparisons were nonsignificant. (C) IFNB1 transcripts were first normalized to β-actin expression and then shown relative to uninfected cells for each siRNA. HRV-A16-infected (MOI: 10) primary human fibroblasts were previously transfected with the indicated siRNA. (D) Similar to C, except for IFNL3 (IL-28) transcripts. (E) Immunoblot showing efficiencies of MAVS and MDA5 knockdown in C and D, except that cells were treated overnight with IFN-α. Data show means ± SD from four independent experiments in C and D, and a representative experiment in E. *, P < 0.05, by Kruskal-Wallis test comparing areas under the curves in C and D.