IFIT5 RNA interference assay

Three small interfering RNAs (siRNAs) targeting the IFIT5 gene were designed and synthesized by GenePharma (Table 2). DF-1 cells were cultured in six-well plates overnight, and 2 μl of Lipofectamine RNAiMAX (Invitrogen) was used to transfect 1 μg of 200 nM IFIT5 of each siRNA separately or the negative control (siNC) to inhibit IFIT5 protein expression. Cell samples and culture supernatants were collected 24 h after transfection and infected with ARV strain S1133 (MOI = 1). We extracted RNA from the cells to detect ARV replication and changes in the expression of molecules related to innate immune signaling pathways. Viral replication was measured in the culture supernatant.

siRNA sequences targeting the IFIT5 gene.