Extended Data Fig. 5: Trans-Tango labeling in stable transgenic lines.

Robust mApple-CAAX labeling in Tg(isl2b.2:Gal4, myl7:tagRFP) 6 dpf larvae bearing both the ligand and arrestin–TEV stable transgenes and Tg(UAS:GFP) and Tg(QUAS:mApple-CAAX) reporter lines. a,a′, Control larvae from injections of plasmids for all trans-Tango components (n = 10). b,b′, Tg(10xUAS-sGCG-ICAM(1235)-nrxn1b, cryaa:mCherry) larvae injected with the receptor and arrestin–TEV constructs (n = 10). c,c′, Tg(elavl3:hArr:TEV, he1.1:YFP) larvae that had been injected with the receptor and ligand constructs (n = 10). d,d′, Larvae transgenic for both the ligand and arrestin–TEV transgenes that had been injected with the trans-Tango receptor construct (n = 10). e, Uninjected controls (n = 10). f, On average, 45 neurons were labeled with mApple-CAAX in larvae that bore both the ligand and arrestin–TEV stable transgenes compared to those from injections of all trans-Tango components (on average, 11 neurons per sample). In the boxplots the center line represents the median, the box limits indicate top and bottom quartiles, and the whiskers extend to the 1.5× quartile range. g, Image registration of mApple-CAAX-labeled neurons in the optic tectum and hindbrain of ten Tg(isl2b.2:Gal4-VP16, myl7:tagRFP); Tg(UAS:GFP); Tg(10xUAS-sGCG-ICAM(1235)-nrxn1b, cryaa:mCherry); Tg(elavl3:hArr:TEV, he1.1:YFP) larvae injected with the receptor construct. mApple-CAAX-labeled neurons are pseudocolored differently for each larva and dashed circles indicate some cells that were labeled in multiple samples. Scale bars, 20 μm. Double asterisk (**) represents the statistically significant increase (P < 0.0001) in trans-Tango labeled neurons when using the ligand and arrestin stable lines compared to plasmid only and stable lines individually using a one-way ANOVA test on Prism.