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Fig. S11 (A-E) Injection of klf2a mismatch morpholino does not affect valve invagination and cell shape. (F) Overexpression of klf2a mRNA rescues klf2a MO-mediated phenotype. (A-B′) Comparison of the valve phenotype between the different treatment affecting valvulogenesis using Tg (flk1:egfp) at 72 hpf. GFP is expressed in the endothelial cell layer and highlights the developing valves. (A, B, and B′) control embryo, (C, D, and D′) klf2a mismatch morphant. (B′ and D′) Schematic representation of the panels (B and D) outlining the endothelial cells within valve-forming region (yellow) and the heart lumen (white). Mismatch MO injection leads to a normal ingression of the endothelial cells and cuboidal cell rearrangement showing that leaflet invagination occurs properly and that there is no nonspecific effects due to MO injection. (F) Percentage of rescue obtained after overexpression of klf2a mRNA concomitantly with klf2a MO (n = 115 for MO1, n = 49 for MO2) compared with klf2a MO injected embryos (n = 84 for MO1 and n = 88 for MO2). A, atrium; V, ventricle.