Drosophila enhancer adult bristle expression assay using UAS sv-RNAi

Data from manuscript "Sex-specific evolution of a Drosophila sensory system via interacting cis- and trans-regulatory changes" by D Luecke, G Rice, and A Kopp.

This study examines the regulation of expression for the chemosensory bristle specifier gene Pox neuro, which has expanded expression into ancestrally mechanosensory bristles in male D. prolongata. Flourescent assays show qualitativetly expanded expression driven by the D. prolongata allele relative to enhancers from sister species in the developing pupal leg. Adult bristle disruption assays using shavenoid RNAi show quantitative expression expansion by the D. prolongata allele into specific mechanosensory bristles on the dorso-anterior leg of the first tarsi.

Methods
Pox neuro enhacer regions were cloned into Gal4 reporter vector pBPGUw and inserted site-specifically into the attP2 docking site. Homozygous reporter-Gal4 flies were crossed to UAS-dcr2; UAS-svRNAi flies. T1 legs from the F1 of this cross were mounted in PVA clearing media and examined under light microscopy. Individual bristles were scored as 0 for missing, 0.5 for truncated, and 1 for intact. Comparisons between number of effected bristle per species enhancer allele were done by pairwise t test, with bonferonni correction for multiple tests.