Figure 4. Loss of MDA5 function results in increased replication of HRV in respiratory epithelial cells. (A) HRV transcripts, normalized to nonspecific siRNA negative (siNeg) control at 20 h. HRV-B14–infected (MOI: 1) A549 cells were previously transfected with the indicated siRNA. (B) Immunoblot showing efficiencies of MDA5 and RIG-I knockdown in A. Transfected cells were left uninfected or infected with HRV-B14 for 48 h. 120 µg of lysates were run per lane. (C) Similar to A, at 48 h after infection and two rounds of transient transfection with the indicated siRNA. (D) Immunoblot showing efficiencies of MAVS and MDA5 knockdown in C. (E) HRV transcripts and HRV simultaneously quantitated by infectious plaque assay, at 60 h after HRV-B14 infection. (F) Number of HRV reads in RNA-seq data during HRV-B14 infection. A549 cells were previously transfected with MDA5 or nonspecific negative control siRNA (siNeg). Mean of triplicate expression ratios from triplicate infections were shown for each time point. (G) HRV transcripts, normalized to nonspecific siRNA negative (siNeg) control at 48 h, HRV-A16–infected (MOI: 10). Human primary fibroblasts were previously transfected with the indicated siRNA. (H) Immunoblot showing efficiencies of MDA5 and MAVS knockdown in G, except that cells were treated overnight with IFN-α. Data show means ± SD from six (A); eight (C), four (E), and seven experiments (G). Representative experiments are shown in B, D, and H. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, by Kruskal-Wallis test (A, C, and G); Mann-Whitney U test in (E); and Kolmogorov-Smirnov test (F); other comparisons in A and F were nonsignificant.
Figure 4.

Loss of MDA5 function results in increased replication of HRV in respiratory epithelial cells. (A) HRV transcripts, normalized to nonspecific siRNA negative (siNeg) control at 20 h. HRV-B14–infected (MOI: 1) A549 cells were previously transfected with the indicated siRNA. (B) Immunoblot showing efficiencies of MDA5 and RIG-I knockdown in A. Transfected cells were left uninfected or infected with HRV-B14 for 48 h. 120 µg of lysates were run per lane. (C) Similar to A, at 48 h after infection and two rounds of transient transfection with the indicated siRNA. (D) Immunoblot showing efficiencies of MAVS and MDA5 knockdown in C. (E) HRV transcripts and HRV simultaneously quantitated by infectious plaque assay, at 60 h after HRV-B14 infection. (F) Number of HRV reads in RNA-seq data during HRV-B14 infection. A549 cells were previously transfected with MDA5 or nonspecific negative control siRNA (siNeg). Mean of triplicate expression ratios from triplicate infections were shown for each time point. (G) HRV transcripts, normalized to nonspecific siRNA negative (siNeg) control at 48 h, HRV-A16–infected (MOI: 10). Human primary fibroblasts were previously transfected with the indicated siRNA. (H) Immunoblot showing efficiencies of MDA5 and MAVS knockdown in G, except that cells were treated overnight with IFN-α. Data show means ± SD from six (A); eight (C), four (E), and seven experiments (G). Representative experiments are shown in B, D, and H. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, by Kruskal-Wallis test (A, C, and G); Mann-Whitney U test in (E); and Kolmogorov-Smirnov test (F); other comparisons in A and F were nonsignificant.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal