Fig. 6: vsiR-1 and vsiR-2 inhibited ZIKV replication in vitro. | Nature Communications

Fig. 6: vsiR-1 and vsiR-2 inhibited ZIKV replication in vitro.

From: Identification of antiviral RNAi regulators, ILF3/DHX9, recruit at ZIKV stem loop B to protect against ZIKV induced microcephaly

Fig. 6

In ZIKV-infected SK-N-BE(2) cells (MOI = 1, 48 hpi), RT-qPCR, western blot, and virus plaque assay analysis of viral copies (A), ZIKV-E, GAPDH expressions (B) and viral titer (C) by scramble or indicated vsiRNA mimic transfection (MOI = 1, 48 hpi). Bars, mean ± SD, n = 3 biological replicates/group, two-tailed Student’s t-test. **p < 0.01, ***p ≤ 0.001. D Western blot showing ZIKV-E protein expression by depletion of DICER or AGO2 in ZIKV-infected control or ILF3- overexpressed cells (MOI = 1, 48 hpi). n = 2 biological replicates/group. E RT-PCR showing viral RNA copies in ZIKV-infected control or ILF3 overexpressed cells with or without DICER or AGO2 presence. Bars, mean ± SD, n = 3 biological replicates/group, two-tailed Student’s t-test. n.s., non-significance, *p < 0.05. In ZIKV-infected control or ILF3-knockdown cells (MOI = 1, 48 hpi), RT-qPCR (F) and WB (G) analysis of viral RNA copies and relative protein expressions by scramble or indicated vsiRNA mimic transfection. Bars, mean ± SD, n = 3 biological replicates/group, two-tailed Student’s t-test. n.s., non-significance, ***p ≤ 0.001. H, I In ZIKV-infected control or DHX9 knockdown cells (MOI = 1, 48 hpi), RT-qPCR analysis (G) and WB (H) analysis of viral RNA copies and relative protein expressions by scramble or indicated vsiRNA mimic transfection. Bars, mean ± SD, n = 3 biological replicates/group, two-tailed Student’s t-test. n.s., non-significance, **p < 0.01, ***p ≤ 0.001. J In vitro NS5 polymerase activity assay measured by control or vsiR-1 mimic treatment. Relative intensity of newly synthesized biotin-labeled RNAs was quantified by Image J software. Bars, mean ± SD, n = 3 biological replicates/group, two-tailed Student’s t-test. **p < 0.01. In vitro (K) and in vivo (L) Luciferase activity of ZIKV 5R3 RNA by control or vsiR-1 mimic transfection. Bars, mean ± SD, n = 3 biological replicates/group, two-tailed Student’s t-test. **p < 0.01. Source data are provided as a Source Data file.

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