Inhibiting hemodynamic flow leads to reduced cardiac Klf2a and Notch signaling.
( A–D) Confocal imaging performed on klf2a:H2B-GFP ; vmhc:mCherry-NTR uninjured hearts at 24 hpt (6 dpf) reveals decreased klf2a:H2B-GFP expression in ( B) blebbistatin (Blebb) and ( C) tricaine (Tric) treated hearts as well as in ( D) gata2a-/- mutant hearts when compared to ( A) ethanol (EtOH)-treated control hearts (n = 12 control EtOh; 6 control Blebb; 7 control Tric; 9 control gata2a-/-). ( E–H) Confocal imaging performed on Tp1:d2GFP ; vmhc:mCherry-NTR uninjured hearts at 24 hpt (6 dpf) shows decreased Tp1:d2GFP Notch reporter expression in hearts treated with ( F) blebbistatin or ( G) tricaine when compared to ( E) ethanol-treated control hearts; however, ( H) gata2a-/-hearts do not display significant changes as quantitatively assessed in Figure 3N(n = 12 control EtOh; 5 control Blebb; 7 control Tric; 9 control gata2a-/-). ( I–L) Whole-mount in situ hybridizations reveal decreased notch1b expression in hearts treated with ( J) blebbistatin (n = 0/9) or ( K) tricaine (n = 0/5) and in ( L) gata2a-/- hearts (n = 4/12) when compared to ( I) ethanol-treated control hearts (n = 10/10) at 24 hpt (6 dpf). All confocal images shown are maximum intensity projections. V, ventricle; A, atrium; dpf, days post-fertilization; hpt, hours post-DMSO treatment. Dashed lines outline the heart. Bars: 50 μm.
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