A novel reciprocal paracrine signaling loop mediated by transient expression of EphrinB2 in the myocardium regulates ventricular wall formation.

Genome-wide gene expression studies on EphB4 mutant over wild-type mice. We used microarray to show enrichment of neuregulin-1 pathway in EphB4 mutant mice A total of 6 mice were used, including 3 EphB4 mutant and 3 wild-type. Total RNA was isolated using Qiagen RNeasy kit (Qiagen, Valencia, CA) from mouse heart. Ten micrograms of total RNA was used to synthesize cDNA. T7 promoter introduced during the first strand synthesis was then used to direct cRNA synthesis, which was labeled with biotinylated deoxynucleotide triphosphate, following the manufacturer's protocol (Affymetrix, San Diego, CA). After fragmentation, the biotinylated cRNA was hybridized to Mouse 430.2 Array chips (Affymetrix, San Diego, CA)