Testing of the unknome set of genes for roles in fertility and wing growth.

(A) Plot of brood sizes obtained from matings in which each gene was knocked down in either the male or female germline. Dotted lines indicate outlier boundaries, with the genes named being those whose position outside of the boundary is statistically significant, error bars show standard deviation, and the size of the circles is inversely proportional to the p-value. Controls: Vret is involved in piRNA biogenesis and affects female fertility [113], and Ref1 is an essential protein predicted to be involved in RNA export [114], and affects both males and females. (B) Summary of the significant hits from the test of male fertility, showing the human ortholog and the phenotype reported for patients with loss of function mutations (PCD, MMAF). (C) Adult wing illustrating the posterior domain that expresses engrailed during development and hence the engrailed-Gal4 driver used to express the hairpin RNAs. Also shown are the intervein areas measured to assess tissue growth in the anterior and posterior halves of the wing. (D) Plot of the mean area of the anterior and posterior intervein areas as in (C) for flies in which each gene was knocked down by RNAi in the posterior domain (pixel dimensions 2.5 μm × 2.5 μm). Errors are shown as tilted ellipses with the major/minor axes being the square roots of the eigenvectors of the covariance matrix. Dotted lines indicate the outlier boundary, with the genes named being those whose position outside of the boundary is statistically significant, with the size of the circles being inversely proportional to the p-value. The genes Hippo (growth repressor) and Chico (growth stimulator) were included as controls. (E) Representative wings from flies expressing hairpin RNA for the indicated genes in the posterior domain. Hippo and Chico are controls as in (D), with CG11103 and CG5885 showing an increase or decrease in the posterior domain, respectively. The means and variances used for the graphs shown in the figure can be found in S2 Data with the data points in S3 Data. MMAF, multiple morphological abnormalities of the sperm flagella; PCD, primary ciliary dyskinesia; RNAi, RNA interference.