(A) Alleles, chromosomal deficiencies, and enhancer trap or T2A GAL4s used in this study, as well as approximate location for primers (F and R) used to validate mutants. (B) Class III expression of subdued and CG15270 from cell-type-specific microarray (GSE69353), expressed as mean log fold-change difference between isolated CIII and whole-larval samples. Positive values indicate enrichment/upregulation and negative values downregulation. (C) qRT-PCR validating CIII expression of subdued and CG15270 (n=4, each condition). (D) UAS-GFP driven under the control of a subdued enhancer trap, GAL4c240 or T2A-GAL4wwk, evidences that subdued and white walker are expressed in CIII neurons. (E) Species, cladogram, and number of sequences used in phylogenetic analysis. (F) Maximum likelihood phylogeny of animal anoctamins, with weak branches rearranged in a species-aware manner against cladogram in E. subdued is a member of the ANO1/2 clade of nephrozoan calcium-activated chloride channels and of the clade of metazoan anoctamins that includes mammalian ANO1/2/3/4/5/6/9. Mammalian ANO8 is homologous to CG15270. Additionally, this phylogeny evidences a separate clade of ANOs of unknown function we have deemed ANO11 (cyan).