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Fig. 4 Comparable vascularization and activation of FGF signaling in regenerating hearts of young and old fish.
(A-D) Fluorescent images of fli1:EGFP signals of regenerating young (A,C) and old (B,D) fish hearts at 7dpa (A,B) and 14 dpa (C,D). Arrowheads point to the fli1:EGFP signals in the regenerating area. DAPI was used for counterstaining. In C and D, not all signals were labeled for simplicity. (E-F′) fli1: EGFP signals in the uninjured heart (E,E′) and in regenerating heart (F,F′) at 14 dpa in young fish. The signals were detected as small clusters throughout the heart. In the regenerating area, the fli1: EGFP signals formed larger clusters than those in uninjured area. E′ and F′ are close up images of the boxed areas in E and F, respectively. Scale bar: 50 μm; the degree of zoom: ×2. (G) Degree of vascularized areas in the regenerating and uninjured areas. The degree was quantified by the ratio of fli1:EGFP signal-positive area in the regenerating area and the uninjured area from single confocal plane. The p-values between young and old fish are shown. (H-I′) Immunofluorescence images of pErk1/2 (magenta) and MHC (green). Closed and open arrowheads point to the pErk1/2 positive signals at the surface of the heart and inside the regenerating area, respectively. H′ and I′ show close ups of the boxed areas in H and I. Scale bar: 50 μm; the degree of zoom: ×4. (J-K′) Immunofluorescence images of mkp3: EGFP (magenta) and MHC (green). Arrowheads point to the mkp3: EGFP signal positive signals at the surface of the heart and inside the regenerating area, respectively. J′ and K′ show close ups of the boxed areas in J and K. Dotted lines indicate the amputated planes. Scale bar: 50μm; the degree of zoom: ×4.
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