Fig 3. Effects of silencing exo-siRNA, miRNA and piRNA pathway key proteins on different orthobunyavirus infections in Aag2 cells.

Silencing of key transcripts by transfection of sequence-specific dsRNA (Ago1, Ago2, Piwi4 or eGFP as negative control) into Aag2 cells, followed by infection at MOI 0.01 at 24 hours p.t.; BUNV-NL (A), SBV-NL (B) or CVV, SATV (C). Cells were lysed at 48 hours p.i. and luciferase activity determined (A, B) or RNA isolated, followed by cDNA synthesis and virus specific qRT-PCR (C). Knockdown of RNAi transcripts was verified by qRT-PCR (D). S7 was used as internal control in qRT-PCR assays. All data were normalized to cells transfected with control dsRNA. Graphs represent three independent experiments performed in triplicate for the luciferase assays and three independent experiments for the qRT-PCR assays. * p≤0.05 student t-test.