Additional file 1 of Monoallelic knockout of r2d2 affects the antiviral RNAi response to Mayaro virus and the reproductive potential in Aedes aegypti

Additional file 1: Figure S1: ECFP eye marker expression in ∆R2D2(+/-) L2/L3 (G1) larvae of Aedes aegypti (M2, blood feed #1; see Table 1) resulting from male G0 survivors (harboring the “ΔR2D2-U6” donor plasmid), which had been backcrossed to the HWE recipient strain of Ae. aegypti. Figure S2: Effects of monoallelic r2d2 KO on mosquito fecundity and fertility. A Fecundity over two gonotrophic cycles of individual ∆R2D2(+/-) and HWE females (n = 20) when outcrossed or intercrossed. Each data point represents the number of eggs per gonotrophic cycle produced by a single female. B Fertility over two gonotrophic cycles of individual ∆R2D2(+/-) and HWE females (n = 20) when outcrossed or intercrossed. Each data point represents the relative quantity of pupae developed from all eggs a single female produced during the first or second gonotrophic cycle. Bars represent mean values. *p < 0.05; **p < 0.01; ***p < 0.0001; Mann-Whitney U-test. Figure S3: Relative dcr2 expression levels in MAYV-infected ∆R2D2(+/-) and HWE females at 4 hpi, 4 dpi, and 10 dpi (virus titer in the blood meal: ~ 107 PFU/ml). Each data point represents a whole-body female. Expression levels of dcr2 were normalized to the average expression of rsp7. Bars indicate median values. Statistical test: Mann-Whitney U-test. Figure S4: Relative mCherry expression levels in MAYV-infected ∆R2D2(+/-) females as analyzed by RT-qPCR A at 4 dpi with MAYV; B at 4 hpi, 4 dpi, and 10 dpi with MAYV. Each data point represents a whole-body female that had acquired a MAYV-containing blood meal (titer in the blood meal: ~ 107 PFU/ml) or a similar blood meal containing no virus. mCherry expression levels were normalized to the average expression of rsp7. Bars indicate median values. ***p = 0.0010; Mann-Whitney U-test. Supplemental Table S1: List of oligonucleotide primers used for the study.