A Doxycycline (monohydrate) Protocol domain (Fig. 6a, b, Supplementary Fig. 6A). In contrast, TaoRNAi in A08n neurons resulted in a strong enhance in C3da 08n SybGRASP puncta, initial detectable at 48 h AEL. No Syb-GRASP puncta had been detected involving A08n and also the a lot more dorso-laterally positioned cho neurons s suggesting that Tao function is essential to restrict C3da 08n synapse formation. To additional test if C3da 08n synapses had been functional, we performed in vivo calcium imaging experiments. We activated C3dacho neurons (nompC-LexA) employing the red-shiftedNATURE COMMUNICATIONS | (2019)10:3506 | 41467-019-11408-1 | www.nature.comnaturecommunicationsNATURE COMMUNICATIONS | 41467-019-11408-ARTICLEUAS-TaoRNAiaC4da Nafcillin Cancer presynapsesControl nsUAS-TaoRNAi ns nsb250 200 150 100 50A08n postsynapsesControlcC4da-A08n synapsesControlUAS-TaoRNAi ns0 48 h 72 h 96 h 120 h 27H06-LexA LexAop-Brpshort-mCherry; 82E12-Gal4UAS-Drep2-GFP0 48 h 72 h 96 h 120 h 48 h 72 h 96 h 120 h 27H06-LexALexAop-Brpshort-mCherry; 82E12-Gal4 UAS-Drep2-GFP 27H06-LexALexAop-Brpshort-mCherry; 82E12-Gal4 UAS-Drep2-GFPdC4daA08n synapses C4da presynapsesControl 80 60 40 20 0 48 h 72 hUAS-TaoRNAi nseC4daA08n synapses A08n postsynapsesControl nsEctopic A08n postsynapsesUAS-TaoRNAi nsf10 eight 6 4 two 0 48 hControlUAS-TaoRNAi80 60 40 20 0 48 h 72 h 96 h 120 h96 h120 h72 h96 h120 h27H06-LexA LexAop-Brpshort-mCherry; 82E12-Gal4 UAS-Drep2-GFP27H06-LexALexAop-Brpshort-mCherry; 82E12-Gal4UAS-Drep2-GFP A08n27H06-LexALexAop-Brpshort-mCherry; 82E12-Gal4 UAS-Drep2-GFPg27H06-LexALexAop-brp-sh-mCherry; 82E12-Gal4UAS-Drep2-GFPC234daC4dahfas3-Cy5 Brpshort-mCherry Drep2-GFPfas3-CyBrpshort-mCherryDrep2-GFPMergeMerge Lateral MedialzUAS-TaoRNAiUAS-TaoRNAiz 27H06-LexALexAop-brp-sh-mCherry; 82E12-Gal4 UAS-Drep2-GFPFig. 4 Tao restricts postsynaptic development and connectivity throughout larval development. a Quantitative evaluation of C4da 08 neurons synaptic profiles from 48 h AEL to 120 h AEL in manage or with TaoRNAi expression in A08n neurons. a C4da neuron presynapse, b A08n postsynapse, and c C4da 08n synapse numbers. Manage 48 h n = 10, 72 h n = eight, 96 h n = 9, 120 h n = ten. UAS-TaoRNAi 48 h n = 10, 72 h n = 9, 96 h n = ten, 120 h n = 8. d C4da 08 synapse to C4da presynapse ratios (in %) through development from 48 h AEL to 120 h AEL in manage or with TaoRNAi expression in A08n neurons. Handle 48 h n = 10, 72 h n = eight, 96 h n = 9, 120 h n = 10. UAS-TaoRNAi 48 h n = 10, 72 h n = 9, 96 h n = 10, 120 h n = eight. e C4da 08n synapse to A08n postsynapse ratios (in %) for the duration of development from 48 h AEL to 120 h AEL in control or with TaoRNAi expression in A08n neurons. Control 48 h n = 10, 72 h n = eight, 96 h n = 9, 120 h n = ten. UAS-TaoRNAi 48 h n = ten, 72 h n = 9, 96 h n = ten, 120 h n = eight. f Quantification of ectopic A08n postsynapses in C2daC3da domain throughout development from 48 h AEL till 120 h AEL. Control 48 h n = 10, 72 h n = 8, 96 h n = 9, 120 h n = ten. UAS-TaoRNAi 48 h n = ten, 72 h n = 9, 96 h n = ten, 120 h n = eight. g Confocal images of hemisegments in manage or with TaoRNAi expression in A08n neurons. Synaptic markers labeling C4da presynapses (magenta), A08n postsynapses (green), and anti-Fas3 immunolabeling C2da, C3da, and C4da sensory axons (blue) at 96 h AEL. Yellow arrowheads show ectopic A08n postsynapses within the C2daC3da domain under TaoRNAi situations. h XZ projections of each and every channel in g are shown. Scale bar = 5 . For all statistical tests: P 0.05, P 0.005, P 0.001, P 0.0001 SD, unpaired two-tailed t-test. For precise P values and.