Extended Data Fig. 3: Analysis of wiring defects in L5 with or without disruption of Ecdysone-pathway TFs.

a, Different L5 arborization defects and their distributions under the given genotypes (see Fig. 2g). All transgenes are expressed using an L5-specific driver (pan-lamina driver is used for data in Fig. 2g). n = number of neurons. **, p-value < 0.001. b, schematic showing the EcR^FlpStop allele. FlpStop cassette is inserted in the first common intron of EcR (grey bar shows insertion site, Mi{MIC}EcRMI05320). Cells expressing Gal4, within which a stochastic, heat-shock FLP recombinase-mediated flipping of the cassette occurs, express tdTom. c, Ca²⁺ response from Tm3 (measured using GCaMP6s) upon optogenetic stimulation of L5 with (blue line) or without (red line) EcR^DN expression in L5. GCaMP6s responses were measured in 3 regions of interest (ROI). ROI 1, ROI 2 and ROI 3 span medulla layers M1, M5 and M10 respectively. (WT, 19 animals; EcR^DN, 17 animals.) d, Ca²⁺ response from Dm13 and Dm18 (common LexA driver used yields expression in both Dm13 and Dm18, see Supplementary Table 1) upon optogenetic stimulation of L5 with (blue line) or without (red line) EcR^DN expression in L5. ROI 1 spans M1 and measures response from Dm18. ROI 2 spans M5 and measures response from Dm13. (WT, 19 animals; EcR^DN, 17 animals.) Note: weak response from Dm18 upon stimulation of L5 irrespective of condition. c, d, Amplitude of relative peak response for each condition is quantified. There is no significant difference between WT and EcR^DN for any comparison shown here. For Dm13 response, see Fig. 2j. For all box-plots, solid line depicts median, while the upper and lower bounds of the box depict the third and first quantile of the data spread respectively. p-value from two-sided Student’s t-test. For number of replicates and exact p-values, see Supplementary Table 10.