Sf-SR-C acts as the receptor for Vip3Aa <i>ex vivo</i>.

Abstract

(A) qRT-PCR analysis of the relative transcript levels of SR genes in the cell lines of Sf9, Sf-pIZT, Sf-SRi1 and Sf-SRi2; Data are expressed as the mean ± SD from three independent experiments; ns, non-significant; *** P < 0.001; one-way ANOVA with Dunnett’s method. (B) Cell mortality of different cell lines (Sf9, Sf-pIZT, Sf-SRi1 and Sf-SRi2) exposed to 50 μg/mL of Vip3Aa for 48 h. Data are expressed as the mean ± SD from three independent experiments; ns, non-significant; ** P < 0.01; one-way ANOVA with Duncan method. (C) Confocal microscopy sections showing the localization of Vip3Aa-RFP (red) and Sf-SR-C (green) in Sf9 cells. The anti-Sf-SR-C-N polyclonal antibody and Alexa Fluor 488-conjugated anti-rabbit antibody were used to show the location of Sf-SR-C in Sf9 cells. Anti-GST polyclonal antibodies were used as the control. Arrowheads point to co-localization between Vip3Aa-RFP and Sf-SR-C. Nuclei are stained with DAPI (blue). Scale bar, 10 μm. (D) S2 cells were transfected with Sf-SR-C or Sf-S2. 48 h after transfection, cells were exposed to Vip3Aa-RFP or RFP (red), fixed, and then immunostained with Dylight 488-conjugated anti-V5 antibodies (green). Nuclei are stained with DAPI (blue). Scale bar, 10 μm. (E) Cell viability of different S2 cell lines (S2, S2-Sf-S2 and S2-Sf-SR-C) exposed to 25 μg/mL of Vip3Aa for 48 h (The transfection efficiency was about 30%). Data are expressed as the mean ± SD from three independent experiments; ns, non-significant; ** P < 0.01; one-way ANOVA with Duncan method.</p