Extended Data Fig. 2: Analysis of morphology and DIP-β expression with or without EcRDN expression.
From: A global timing mechanism regulates cell-type-specific wiring programmes
a, Morphology of lamina neurons (L1-L5) in wildtype (WT) brains and upon pan-lamina expression (using 9B08 Gal4) of EcRDN, EcRDN + EcR-B1 cDNA or EcR RNAi. Arrowhead in inset points to M3 medulla layer. Note: loss of arborization in M3 with EcRDN is likely due to loss of driver expression in L3 neurons (see Extended Data Fig. 2b). b, Morphology of L3 neurons with or without EcRDN expression using an L3-specific driver (9D03 Gal4). Note, in adults 9D03 Gal4 also labels some L2 neurons. This is not the case during development (see Extended Data Fig. 11f, g) a, b, Arrowhead points to M3 layer in the medulla. Scale bar, 50μm. c, Effect of EcRDN expression on morphology of T4/T5 neurons. Four layers in the lobula plate, a, b, c and d, are marked with red, yellow, blue and pink arrowheads, respectively. Cartoons of one T4 (purple neuron, top) and one T5 neuron (purple neuron, bottom) are shown to highlight wildtype morphology. Scale bar, 20 μm. d, DIP-β mRNA expression in L1-L5 neurons across development with or without pan-lamina expression of EcRDN (from scRNA-Seq based transcriptomic analyses, see Fig. 3, Extended Data Fig. 4, 5). * significant difference between control and EcRDN (change in expression > 2-fold, p-value < 0.05, two-sided Wilcoxon rank sum test). e, Staining using anti-DIP-β antibody at the indicated time points in development with or without pan lamina expression of EcRDN. Inset shows the lamina neuropil. Arrowhead points to proximal lamina neuropil, positive for DIP-β staining at 72hAPF. Note that staining is largely absent from the lamina neuropil (yellow arrowheads). The level of DIP-β RNA is reduced in EcRDN at 48 but rises to near normal levels again at 72 h. We assume that the decrease in RNA leads to a lag in the accumulation of DIP-β protein. Scale bar, 50 μm. f, EcR-usp complex binding motif within the first intron of DIP-β (FDR < 10−7, see Methods). For number of replicates and exact p-values, see Supplementary Table 10.
