Effect of egr3 knockout on gene expression of dissected zebrafish hearts
To screen for Egr3 targets mediating cardiac valve development we assessed gene expression on dissected hearts of egr3 mutants and wild-type siblings. Overall design: To evaluate the transcriptional differences underlying the egr3 mutant phenotype, we conducted a bulk RNA-seq analysis in dissected zebrafish hearts at 48 hpf.  We dissected 20 hearts for each biological duplicate of egr3 mutant and wild-type sibling samples in cold DMEM with 10% FBS.  Total RNA was isolated using the miRNeasy micro Kit (Qiagen 217084), followed by on-column DNase digestion (DNase-Free DNase Set, Qiagen 79254), and final elution was performed in 12µl of RNase-free water.   Analysis of the obtained reads confirmed the 11bp deletion in the mutant samples.   1. Boezio, G. L. M. et al. The developing epicardium regulates cardiac chamber morphogenesis by promoting cardiomyocyte growth. Dis Model Mech 16 (2023). https://doi.org:10.1242/dmm.049571