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Supplemental Figure 2. Altering HCN4 expression does not disrupt general axis patterning, germ layer specification, or myocardial differentiation. (A) The expression pattern of Xbra, Otx2, and Nkx2.5 in HCN4-DN(AAA) injected embryos was not different from stage-matched, sibling controls as seen by ISH. Representative images are from uninjected sibling controls and HCN4-DN(AAA) one of 2-cell (left) injected embryos. n=30, N=1 for Xbra (vegetal view) and Otx2 (anterior view, dorsal at top). n=25, N=1 for Nkx2.5 (ventral view anterior at top). dorsal lip of blastopore denoted by a yellow dashed line. (B) Shh expression is also not disrupted with injection of HCN4-OE or HCN4-DN(AAA). At gastrulation, Shh is whisker-shaped above the dorsal lip of the blastopore (black line; vegetal view). As development progresses it is found along the midline, in the notochord and somites. n=30, N=1 for stage 10.5 and stage 21 (dorsal view, anterior at top). n=50, N=1 for stage 29/30 (lateral view, anterior at top). Scale bars=200 µm. |
