To generate LexAop-nompC-L-GFP-2A-tdtomato flies, we subcloned nompC-L His-GFP from pOCC8 nompC-L His-GFP [the plasmid was a gift from Z. Wang (Institute of Neuroscience, China)] using the following primer sequences: 5′-cctttacttcaggcggccgcggcccgcaatgtcgcagccgcgcg-3′ (forward) and 5′-gttggtggcggtaccgctgcctccactgtgatggtgatggtg-3′ (reverse). tdtomato was subcloned from pDEST-HemmarR tdtomato [the plasmid was a gift from Y. Rao (Peking University, China)] using the following primer sequences: 5′-ggcagcggtaccgccaccaacttcagcctgctgaagcaggccggcgatgtggaggagaaccccgggcccatggtgagcaagggcgaggag-3′ (forward) and 5′-gtaaggttccttcacaaagatcctttagagggcaacttcattttc-3′ (reverse). We inserted nompC-L His-GFP and tdtomato into pJFRC19-13XLexAop2-IVS-myrGFP vector and then injected the LexAop-nompC-L-GFP-2A-tdtomato plasmids into the transgenic flies (BDSC no. 25710).

To generate LexAop2-FRT-CsChrimson.mVenus-FRT flies, we subcloned CsChrimson.mVenus from the fly 20XUAS-IVS-CsChrimson.mVenus (BDSC no. 55135) using the following primer sequences [containing the FRT (flipase recognition target) sequence]: 5′-cttatcctttacttcaggcggccgcgaagttcctatactttctagagaataggaacttcgccaccatgagcagactggtcgccgctt-3′ (forward) and 5′-aggttccttcacaaagatcctctagagaagttcctattctctagaaagtataggaacttcttacacctcgttctcgtagcaga-3′ (reverse).

CsChrimson.mVenus was inserted into the Not I/Xba I–digested pJFRC19-13XLexAop2-IVS-myrGFP vector (Addgene no. 26224), and the LexAop2-FRT-CsChrimson.mVenus-FRT plasmids were injected into the transgenic flies (BDSC no. 25710).

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