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Fig. 4 | Phytopathology Research

Fig. 4

From: Gelsolin of insect vectors negatively regulates actin-based tubule motility of plant reoviruses

Fig. 4

Gelsolin negatively regulated Pns11 expression and RGDV accumulation. a RGDV infection up-regulated the expression of gelsolin. (I) RT-qPCR assay showing the relative expression of gelsolin, Pns11, and outer capsid protein P8 in cultured cells at 48 hpi, compared to that at 12 hpi. The expression levels of P8, Pns11, and gelsolin in RGDV-infected cells at 12 hpi were normalized as 1. Error bars indicate standard deviations from three independent RT-qPCRs. (II) Western blots assay showing the expression of gelsolin in uninfected and RGDV-infected cultured cells at 48 hpi. b Knockdown of gelsolin improved the level of Pns11 expression and RGDV accumulation in cultured cells. The expression levels of P8, Pns11, and gelsolin in dsGFP-treated cells were normalized as 1. Error bars indicate standard deviations from three independent RT-qPCRs. A one-tailed t-test was conducted in GraphPad Prism 6. The mean results ± the standard deviations are shown. * P < 0.05. c Knocking down gelsolin expanded the viral infection area and increased Pns11 expression in cultured cells. Cells were immunolabeled, for virus with RGDV -FITC (green) and for Pns11 tubules with Pns11-rhodamine (red), then examined by confocal microscopy. Bars, 10 μm. d Knocking down gelsolin increased viral titer in insects. Nonviruliferous second-instar nymphs sequentially were allowed a 3-day acquisition of RGDV and subjected to dsRNAs microinjection, then more than 30 living insects were examined at 6 days padp. e Knockdown of gelsolin increased the viruliferous rate of N. cincticeps. The dsRNA-microinjected insects, which then fed on RGDV-infected plant for 2 days, were analyzed for the viruliferous rates by RT-PCR at 4-day padp. Error bars indicate standard deviations from three independent RT-qPCRs. Two-tailed t-test analysis was conducted in GraphPad Prism 6. The mean results ± the standard deviations are shown. * P < 0.05

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