Ectopic gsx2 expression alone does not induce IO neurons. (A,B,D,E) Expression of gsx2 in 3 dpf Tg(UAS:gsx2BLRP-P2A-BirA-P2A-mCherry) (control, A,D, n=5) and TgBAC(ptf1a:Gal4-VP16);Tg(UAS:gsx2BLRP-P2A-BirA-P2A-mCherry) (B,E, n=5) larvae. The Tg(UAS:gsx2BLRP-P2A-BirA-P2A-mCherry) line harbors a transgene containing the open reading frame (ORF) of gsx2, biotin ligase recognition peptide (BLRP), 2A peptide sequences of porcine teschovirus-1 (P2A), biotin ligase (BirA) and mCherry genes. Expression of Gsx2 and mCherry can be driven by Gal4-VP16. (C) Expression of mCherry was induced where ptf1a was expressed in 3 dpf Tg larvae. (F,G) Expression of pou4f1 in 3 dpf control (F, n=3) and Tg(ptf1a:Gal4-VP16);Tg(UAS:gsx2BLRP-P2A-BirA-P2A-mCherry) (G, n=3) larvae. Although gsx2 was ectopically expressed in all ptf1a-expressing cells, it did not induce ectopic expression of pou4f1. Scale bars: 100 µm in A (applies to A,B,D-G); 50 µm in C.
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