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. Author manuscript; available in PMC: 2011 Apr 30.
Published in final edited form as: Circ Res. 2010 Mar 25;106(8):1342–1350. doi: 10.1161/CIRCRESAHA.109.213132

FIGURE 4. Sodium channel knockdown perturbs the expression of cardiac precursor genes in anterior lateral mesoderm.

FIGURE 4

A. Analysis of gene expression in the early heart-forming region of anterior lateral plate mesoderm at the 6-somite stage by in situ hybridization. Embryos injected with scn5Laa translation inhibitor morpholino (AA_MO1) had reduced expression of nkx2.5 gata4, and hand2 but no change in gata5 expression compared to embryos injected with 5-mismatch control morpholino (AA_MIS1). All embryos shown are from the same injected clutch. Scale bar = 200µm. B. Real-time quantitative RT-PCR performed using independent clutches of embryos also revealed significantly reduced nkx2.5 expression but no change in gata5 expression following injection of AA_MO1 compared to injection of AA_MIS1 control morpholino. Results are mean ± s.e.m for three independent experiments, each performed in triplicate. P<0.05 for nkx2.5, T-test. C. Quantification of numbers (n) of scn5Laa morphant embryos displaying absent, reduced or normal (comparable to wild type) expression levels of cardiac precursor genes as assessed by in situ hybridization. D. Knockdown of scn5Lab also resulted in markedly-reduced expression of nkx2.5 and gata4 at 6 somites. However, no significant change in the expression of hand2 was observed. E. Sodium channel morpholinos reduce nkx2.5 expression in a dose-dependent manner as shown by RT-PCR (AA_MO1) and in situ hybridization at 6 somites (AB_MO2).