Fig 1.
Overview of experimental design.
In experiment 1, we studied CHIKV and DENV loads in siblings from single female x male pair crosses and calculated the heritabilities and genetic correlations of the loads. We then carried out RNAseq comparing two groups; families with the four highest and four lowest mean CHIKV loads. In experiment 2, we reared a separate set of mosquitoes from the same population and exposed them to CHIKV, DENV, or mock (uninfected cell culture control) infection through blood-feeding treatments and then dissected key tissues (midgut, carcass, salivary glands) at three timepoints post-infection (4, 7, 14). We then examined patterns of gene expression of the top candidate gene from the RNAseq analysis in experiment 1 across treatments and tissues x time. Created with BioRender.com.
Fig 2.
Ranked families by viral load.
We ultimately obtained data from 37 families from >600 initial paired crosses. A) DENV load-ranked families and B) CHIKV load-ranked families. Each point represents a single infected mosquito. Box plots show mean viral load ± SEM of each distinct Ae. aegypti family. Whole mosquito viral load was log10 transformed. N = 3–5 individuals per family.
Fig 3.
Scatter plot showing the relationship between mean DENV and CHIKV loads from individual families.
Gray area represents the 95% confidence interval of the regression line. N = 37 mosquito families.
Fig 4.
RNAseq counts for the two loci with significant expression differences between high and low CHIKV-load families.
Points depict normalized read counts of individual families, bars show group medians, and shaded area demarcates the range for each group. Significance was evaluated using Wald tests while correcting for multiple comparisons.
Fig 5.
Tissue-specific expression of AAEL004181 across time in (A) CHIKV- and (B) DENV-infected mosquitoes compared to expression in mock controls. Points depict relative expression level of AAEL004181 in individual samples, while horizontal lines mark group medians. Significance was evaluated by Mann-Whitney U tests between mock and control. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001. Note that a day 7 salivary gland outlier in the mock treatment group has been omitted from both panels (value = 7.618). dpi, days post-infection. n = 20 per treatment.
Table 1.
Primers and probes for virus quantification.