PMC

(A) The phenotypes of nix⁺ females. Thirty individuals were sacrificed at the larval stage to examine gene expression and therefore have an undetermined phenotype. (B) The number of nix⁺ females with male-specific features from experiment 1. (C) Wild-type genitals compared with the genitals of a nix⁺ female, which have gonocoxites and gonostyli. (D) Wild-type testes and ovaries compared with gonads of a nix⁺ female, which had testes and accessory glands. Wild-type images and nix⁺ images are viewed under 55× and 80× magnification, respectively. AG, accessory glands; VD, vas deferens.

(A) Phenotypes of injected individuals. (B) The female isoform of dsx (dsx^F) is present in nix⁻ A. aegypti detected by ddPCR. Rel. Exp., relative expression. Error bars, mean ± SEM. (C) The female isoform of fru (fru^F) is present in the nix⁻ A. aegypti detected by ddPCR. Error bars, mean ± SEM. (D) Feminization of the antennae in a nix⁻ male individual. (E) Feminization in the genitals of a nix⁻ male individual. (F and G) The log2 reads per kilobase per million mapped reads expression level heat map of the top 100 male-biased (F) and female-biased (G) genes in wild-type males, nix⁻ males, and wild-type females. Two heat maps from nix⁻ males are shown here. All other heat maps are shown in .

(A) PCR for Nix in male and female genomic DNA. For all PCR experiments, a ribosomal protein gene, RPS7, was used as a positive control. (B) PCR for Nix in genomic DNA from recombinant female m^J2sensor/m A. aegypti. (C) Reverse transcription PCR expression profile of Nix from 0- to 12-hour embryo cDNA starting at 0 to 1 hour in 1-hour increments. (D) FISH with a probe for Nix and the J2 transgene in mitotic chromosomes of J2 transgenic males. (E) Nix copy number as determined by digital droplet PCR (ddPCR) on male and female genomic DNA. Error bars, mean ± SEM.