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. 2018 Nov 14;3(6):e00549-18. doi: 10.1128/mSphereDirect.00549-18

FIG 4.

FIG 4

Validation of RBM14 as an IAV-required host factor. (A) A549 cells were transfected with three RBM14-targeting siRNAs (siQ1, siQ2, and siQ3). At 48 h posttransfection, the cells were infected with influenza A/WSN/33 (H1N1) virus at an MOI of 0.01 for 24 h. Protein lysates were prepared and subjected to SDS-PAGE and Western blot analysis, immunoblotting for RBM14 (top panel) and the loading control COXIV (bottom panel). (B) RBM14 levels were quantified from the Western blot shown in panel A. (C) A549 cells were transfected with three RBM14 siRNAs (siQ1, siQ2, and siQ3), two siRNAs targeting ARCN1 (siD1 and siD2), the positive-control siNP, or the negative-control siNT. At 48 h posttransfection, the cells were infected with influenza A/WSN/33 (H1N1) virus at an MOI of 0.01 for 24 h. Cellular RNA was extracted, and the expression level of WSN neuraminidase (NA) mRNA was measured by qRT-PCR and normalized to GAPDH in each sample and then to the siNT control. Error bars indicate standard deviations of the means. Data shown represent results of three technical repeats for each of three biological repeats. Statistically significant P values are shown in the figure.