Motile ciliated cells are functionally defective in Wdr92 mutants. (A) Box plot (median and interquartile range) of adult climbing assay for two independent Wdr92 RNAi lines crossed to scaGal4 driver (Table S4). Climbing index (proprioceptive performance) is reduced relative control flies (scaGal4 crossed to KK parent line). n = 5 batches of 10–15 flies. (B) Fertility assay (mean and SD of progeny per male, n = 10 males) for Wdr92 RNAi lines crossed to BamGal4 driver. Controls are males from KK parent line or OregonR (Or-R) crossed to BamGal4. (C) Schematic diagram of generation of Wdr92 deletion mutation by ORF replacement with mini-white gene, mediated by CRISPR/Cas9 using homology-directed repair. (D) Adult climbing assay (median and interquartile range) of Wdr92^−/− deletion mutant, including rescue by Wdr92-mVenus fusion gene. (E) Larval hearing assay. Retraction index (median and interquartile range) reflects the larval retraction response before and during a 1000 Hz tone. n = 9 batches of five larvae. (F) Control male reproductive organs, showing motile sperm within SV and ejaculatory duct (ED), and maturing sperm bundles (SB) in the testes. (G) Wdr92^−/− mutant, showing sperm bundles, but lack of motile sperm in SVs or ED, consistent with sperm motility being required for transfer to these structures (Moore et al., 2013). Significance was determined by Kruskal-Wallis test, with Dunn’s test for multiple comparisons (A, D, and E) or ordinary one-way ANOVA, with Dunnett’s test for multiple comparisons (B). Significance on plots is signified by asterisks: *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001; ****, P ≤ 0.0001. Bars, 100 µm.