File:The-cation-channel-TRPA1-tunes-mosquito-thermotaxis-to-host-temperatures-elife-11750-fig3-figsupp1.jpg

English: (A) Genomic organization and targeted mutagenesis of AaegTRPA1. Non-coding (white) and coding (black) exons are shown to scale (vectorbase.org). Introns are denoted by connecting lines (not to scale). The AaegTRPA1 ZFN targets exon 12 (purple arrow). An insertion cassette (shown to scale), including the Ae. aegypti polyubiquitin (polyUB) promoter driving expression of enhanced cyan fluorescent protein (ECFP) and an SV40 polyadenylation signal, is integrated via homology-directed repair. (B) Schematic of the thermal gradient assay (top, side view). Representative experimental image (bottom, top view) showing mosquitoes outlined in red detected across one lane of the thermal gradient assay, with instantaneous air temperature reported for each of 8 analysis sectors. (C,D) Percent of mosquitoes of indicated genotypes detected in each sector and mean sector air temperature (rounded to nearest °C) in the absence (C) or presence (D) of a thermal gradient (n = 6 trials per genotype). Data are plotted as mean ± s.e.m. (***p < 0.001; two-way ANOVA with Bonferroni correction compared to wild-type). (E) Heat maps showing mean air temperature (left) and percent of mosquitoes of the indicated genotypes detected (right) in each sector over 90 min from the onset of a thermal gradient. (F) Same data as in (E), showing total percent of mosquitoes of the indicated genotypes detected in sectors 1–3 (cold side, solid line, mean; s.e.m., shading) and sectors 6–8 (hot side, dotted line, mean; s.e.m., shading) during onset and maintenance of a thermal gradient. G, Percent of total mosquitoes of the indicated genotypes found dead in sectors 6–8 at the conclusion of the experiment. Each replicate is indicated by a dot, and mean ± s.e.m. by lines. Genotypes with different letters are significantly different (p < 0.01, one-way ANOVA with Bonferroni correction).