Promoter elements have distinct roles in driving hematopoietic, renal and testicular expression of Hemogen in transgenic Tg(Hemgn:mCherry) zebrafish. (A) Schematic of the zebrafish Hemogen gene. CNEs, black; coding exons, white; transcription initiation sites, bent arrows. Three Tg(Hemgn:mCherry,myl7:EGFP) transgenes driven by portions of the Hemogen promoter were transfected into one-cell TU embryos by Tol2 transposase-mediated insertion. Numbers indicate length of promoter elements and arrows show gene direction. (B) 20 hpf. P1 transgene expression in the peripheral blood island (PBI). (C) 72 hpf. P1 transgene expression in the pronephric ducts (PD). (D) 5 dpf. P1 transgene expression in the proximal convoluted tubule (PCT). (E,F) 72 hpf. Co-localization of mCherry and EGFP in progenitors in the CHT of Tg(Hemgn-P1:mCherry,Lcr:GFP) or Tg(Hemgn-P1:mCherry,CD41:EGFP) zebrafish. (G) Transgene expression in mature erythrocytes from adult zebrafish. (H) Transgene expression in adult head kidney (HK), trunk kidney (TK) and tail kidney (T) near the EGFP+ heart (H). (I) Transgene expression in adult Sertoli cells (Se) that surround the seminiferous tubules (ST). (J) Proportion of embryos expressing transgenes P1, P2 or P3 in ICM, kidney, CHT and circulating primitive erythrocytes (RBC). Scale bars: 100 µm (B,D-F,I); 500 µm (C,H); 25 µm (G).