Fig. 4: RNAi knockdown and virus replication in flies.
a Experimental procedure to test antiviral activity of candidate proteins in flies. The fly Gal4/Gal80TS system allows for the temperature-sensitive expression of sh or inverted repeat RNA. Flies carrying both the Gal4/Gal80 and UAS-siRNA were moved to 29 °C, activating Gal4 and inducing the siRNA KD. Upon confirming the viability of the KD flies, they were individually infected via injection of DCV (500 pfu/fly), FHV (500 pfu/fly), CrPV (5 pfu/fly), SINV (2,500 pfu/fly), and VSV (10,000 pfu/fly). Viral replication was measured by RT-qPCR at 2 (CrPV, DCV, FHV) or 3 (SINV, VSV) days post-infection. b Heatmap showing fold change of virus gene expression normalized to the housekeeping gene RP49 upon target protein KD as compared to mCherry KD flies. Data represents the mean of biological triplicates. Significance was calculated using lsmeans R package (least-squares means, linear model) with Dunnet’s adjustment for p values for multiple hypothesis testing. Candidates were further categorized into DNA, RNA, and/or 2′5′OA interacting proteins according to the results of the NA-AP-MS screen. Source data are provided as a Source Data file.
