Fig. 2: dsx^M2 has a potentially masculinizing function like dsx^M.

a RNAi targeting dsx^M and dsx^M2 as indicated by blue and green arrows, respectively. b, c Relative mRNA expression levels of dsx^M (b) and dsx^M2 (c) in control and RNAi-mediated males. n = 9 based on three replicates for each. ***p < 0.001, Mann-Whitney U test. d, e Knocking down dsx^M or dsx^M2 in dsx-expressing cells (d) or pan-neuronally (e) impairs male courtship and mating success with females. n = 23, 24, 24, 24, 24, 24 for dsx^GAL4/+, dsx^GAL4/UAS-dsx^MRNAi, dsx^GAL4/UAS-dsx^M2RNAi, R57C10-GAL4/+, R57C10-GAL4/UAS-dsx^MRNAi and R57C10-GAL4/UAS-dsx^M2RNAi respectively. For courtship index, **p = 0.0012 and ***p < 0.001, Kruskal-Wallis test with Dunn’s multiple comparisons test; for percentage mated, **p = 0.0087 and ***p < 0.001, Chi-square test. f, g Overexpression of Dsx^F feminized, while overexpression of Dsx^M or Dsx^M2 masculinized dsx-expressing cells, including external genitalia and sex comb (arrowhead), under wild-type background [f, dsx^GAL4(∆2)/+] or mutant background [g, dsx^GAL4(∆2)/dsx^1649-9625] for dsx. h, i Relative mRNA expression levels of dsx target genes, yp2 and yp3, in females (h) and males (i). Expression levels of yp2 and yp3 were increased with Dsx^F overexpression and decreased with Dsx^M or Dsx^M2 overexpression. n = 9 based on three replicates for each. ***p < 0.001. Mann-Whitney U test. Error bars indicate SEM.