Figure 1.

fle Is a Sex Determination Gene Necessary for dsx Splicing and Survival in Anopheles Females

(A) A schematic of the A. gambiae fle transcript, with the location of RNAi targets within the coding sequence.

(B) RT-PCR analysis of the dsx splicing pattern in A. gambiae Sua5.1 cells transfected with in vitro-synthesized fle_1 dsRNA, as compared with control non-transfected cells.

(C) Sequences of the putative Fle binding sites in the A. gambiae dsx and fru female-specific exons. Presented are 13-nt fragments with similarity to the Drosophila repeat elements [TC(T/A)(T/A)CAATCAACA].

(D) RT-PCR analysis of fle transcription during A. gambiae development. E, embryos; L, larvae; P, pupae; A, adults; (−), negative control; M, male; F, female. Ribosomal protein S7 (rpS7) transcript levels were used as a gel loading control.

(E and F) Knockdown of fle expression causes female-specific lethality in A. gambiae embryos. A summary of three independent microinjection experiments using fle_1 (E) and fle_2 (F) dsRNA fragments, with bla dsRNA used as control. GFP−/+ denote cohorts of individuals scored during the first larval instar as GFP-negative or GFP-positive. ^∗∗p < 0.0001; Fisher’s exact test.

(G) Knockdown of fle ortholog expression causes female-specific lethality in A. stephensi embryos. A summary of two independent fle dsRNA microinjection experiments. GFP−/+ as in (E). ^∗∗p < 0.0001; Fisher’s exact test.

See also Figures S1–S3.