E2a is necessary for Smad2/3 dependent transcription and the direct repression of lefty

We characterized the binding of Smad2/3 using ChIP-SEQ in both control gastrula-stage X. tropicalis embryos and embryos depleted of the transcription factor E2a (E2a-MO). We also characterized gene expression in control and E2a-depleted embryos by RNA-Seq. For ChIP-Seq, three biological replicates of E2a-MO were performed, and two biological replicates of Control embryos were performed. For RNA-Seq, two biological replicates of E2a-MO were performed, and two biological replicates of Control embryos were performed, and the mean expression levels of each gene were compared. For ChIP-Seq, three biological replicates were performed for E2a-depleted X. tropicalis embryos, and two biological replicates were performed for control gastrula-stage embryos. For RNA-Seq, two biological replicates were performed for both E2a-depleted embryos and control embryos, and the mean expression levels were compared for each gene.