Gene expression signatures for gata4/5/6 knocking down in zebrafish

Zebrafish gata4/5/6 are known to lie on the apex of the regulatory hierarchy in primitive myelopoiesis. However, little is known about the roles of microRNAs in this process. Performing microarray analysis on the expression changes of microRNAs in the gata4/5/6 knockdown embryos, we found that miR-210-5p is a novel regulator in zebrafish primitive myelopoiesis. To uncover the target genes of miR-210-5p to mediate its role in inhibiting zebrafish primitive myelopoiesis, we performed microarray analysis to identify differentially expressed genes in gata4/5/6 knockdown embryos. The embryos microinjected with control MO or gata5/6 MOs at 1-cell stage were collected at 14 hpf for analyzing the gene expression changes affected by knocking down gata4/5/6.The microinjection amount was 1 nl solution containing 6.25 ng gata5 MO and 1.25 ng gata6 MO, or 7.5 ng control MO per embryo. 2 replicates of gata5/6 knockdown embryos and 3 replicates of control embryos were analyzed. Please note that as it is reported that morpholinos against gata5 and gata6 also lead to a loss of gata4 expression in zebrafish (PMID 17869240), a combination microinjection of gata5 and gata6 morpholinos was used for knocking down all the three genes.