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(A) RT-qPCR analysis on sorted Tg(fli1:EGFP)+ versus Tg(fli1:EGFP)− cardiac ventricular cells (uninjured siblings, n = 3; 96 hpci, n = 3). (B and C) Wholemount fluorescence images of (B) (wt siblings, n = 6; mut, n = 6; 96 hpci) Tg(fli1:EGFP) expression in ventricles and quantification of percentage of injured area covered by GFP+ endocardial cells (C). (D) Experimental design and confocal images of immunostaining (mCherry, magenta; αSMA, green) on cryosections from (wt siblings, n = 6; mut, n = 5; 7 dpci) Tg(fli1:CreER); Tg(ubb:GSR) ventricles. (E and F) Quantification of percentage (E) and density (F) of fli1+-derived αSMA+ cells in the injured area, 7 dpci. (G) Schematic and RT-qPCR analysis on sorted Tg(fli1:EGFP)+ cells from (wt siblings, n = 5; mut, n = 3; 96 hpci) ventricles for EndoMT-associated gene mRNA levels. Data represent means ± SD (C, E, and F), and box plots (A and G) show median, IQR (box margins), and 5th and 95th percentiles (whiskers). Student’s t tests (A, C, and E to G). n, pools of two ventricles (A and G), n, ventricles (B to F). Yellow dashed lines demarcate the injured area (B and D); yellow arrowheads point to fli1+-derived αSMA+ cells (D, insets). Ct values are listed in table S5. Scale bars, 200 μm (B), 100 μm (D), and 10 μm (D, insets).
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