The enhanced green fluorescence protein (EGFP) protein expression pattern in the Tg(5' 5kbscl:EGFP)^a transgenic fish. A-H: Lateral (A-C,G,H) and dorsal (D-F) views of the 6-somite (6s), 12-somite (12s), 18-somite (18s), 24 hours postfertilization (hpf), and 48 hpf stage Tg(5' 5kbscl:EGFP)^a embryos, respectively, show the EGFP protein expression in a manner similar to the endogenous Scl protein expression pattern. I,J: Lateral views of 18-somite stage (18s) Tg(5' 5kbscl:EGFP)^a embryos immunohistochemistry stained with anti-EGFP (I, green) and anti-zebrafish Scl (J, red) antibodies. K: Superimposed view of I and J (original magnification, x10). I'-K' Confocal images (x100 magnification) of the boxed region in I, J, and K confirm the colocalization of the EGFP protein (green) with the endogenous Scl protein (red). In all the panels, embryos are oriented with anterior to the left.

The enhanced green fluorescence protein (EGFP) transcript expression pattern in the Tg(5' 5kbscl:EGFP)^a transgenic fish. A-J: Flat-mount (3s, 6s, and 12s) and lateral (18s and 22s) views of the 3-somite (3s), 6-somite (6s), 12-somite (12s), 18-somite (18s) somite, and 22 hours postfertilization (hpf) Tg(5' 5kbscl:EGFP)^a embryos (anterior to the left) show the endogenous scl (A-E) and EGFP (F-J) transcripts in the anterior lateral mesoderm (ALM) and posterior lateral mesoderm (PLM) region. The EGFP transcript in the middle region of the PLM indicated by arrows is drastically reduced in 18s and 22 hpf embryos. K,L: Flat-mount views (original magnification, x10) of the 10s Tg(5' 5kbscl:EGFP)^a embryos (anterior to the left) show colocalization of the EGFP transcript (green) and endogenous scl (red) mRNA. M: Superimposed view of K and L. K'-M': Confocal images (original magnification, x40) of the boxed region in K, L, and M show the EGFP transcript-positive (green) and endogenous scl transcript-positive (red) cells. N,O: Lateral views (original magnification, x10) of the 18s Tg(5' 5kbscl:EGFP)^a embryos (anterior to the left) indicate that the EGFP transcript (green) and endogenous scl mRNA (red). P: Superimposed view of N and O. N'-P': Confocal images (original magnification, x25) of the boxed region in N, O, and P show partial colocalization of the EGFP transcript (green) with the endogenous scl mRNA (red). Q,R: Lateral views (original magnification, x10) of the 20s Tg(5' 5kbscl:EGFP)^a embryos (anterior to the left) indicate that the EGFP transcript (green) and the endogenous embryonic βe1-globin mRNA (red). S: Superimposed view of Q and R. Q'-S': Confocal images (original magnification, x25) of the boxed region in Q, R, and S show that the EGFP transcript (green) is not overlapped with the endogenous embryonic βe1-globin mRNA (red).

The enhanced green fluorescence protein (EGFP) protein is colocalized with embryonic βe1-globin, pu.1, and flk1 transcripts in the Tg(5' 5kbscl:EGFP)^a transgenic fish. A,B: Lateral views of the 20-somite stage (18s) Tg(5' 5kbscl:EGFP)^a embryo stained with the anti-EGFP antibody (green) and antisense embryonic βe1-globin RNA probe (red) show the EGFP protein expression and the endogenous embryonic βe1-globin transcript. A',B<',A'/B': Confocal images (original magnification, x25) of the boxed region in A and B indicate that the EGFP protein (green) is colocalized with embryonic βe1-globin transcript (red). C,D: Lateral views of the 18-somite stage (18s) Tg(5' 5kbscl:EGFP)^a embryo stained with the anti-EGFP antibody (green) and antisense pu.1 RNA probe (red) show the EGFP protein and pu.1 RNA expression. C',D',C'/D': Confocal images (original magnification, x50) of the boxed region in C and D confirm that the EGFP protein (green) is colocalized with pu.1 (red). E,F: Lateral views of the 22 hours postfertilization (hpf) stage Tg(5' 5kbscl:EGFP)^a embryo stained with the anti-EGFP antibody (green) and antisense flk1 RNA probe (red) indicate the EGFP protein and flk1 RNA expression. E',F',E'/F': Confocal images (original magnification, x25) of the boxed region in E and F show the colocalization of the EGFP protein (green) and flk1 (red). In all the panels, embryos are oriented with anterior to the left.

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