Hematopoietic stem cells (HSCs) are capable of self-renewal and differentiation into all mature hematopoietic lineages. This is regulated by transcription factors and chromatin factors to orchestrate chromatin structure and establish an epigenetic code that facilitates gene expression. To uncover chromatin factors that are necessary for the establishment of HSCs, we conducted a reverse genetic screen in the zebrafish using morpholinos to target 488 chromatin remodeling factors. We identified 29 genes that alter HSC marker expression, c-myb and runx1, upon knockdown. Several components of chromatin remodeling complexes already known to regulate hematopoiesis were required for HSC development in our screen. We also found components from complexes that have no known role in HSCs, including the iSwi and Hbo1 complexes. Four members of the Hbo1 complex, Ing4, Phf16, Hbo1, and Brd1, show loss of HSC marker expression upon knockdown. This complex regulates gene expression through the binding of H3K4me3 by Ing4, which results in localized histone acetylation by Hbo1. Ing4 has also been shown to negatively regulate the transcription factor NF-kB through sequestration of the RelA component of NF-kB. To test the function of the complex in HSCs, we used low doses of morpholinos to partially inhibit expression of each complex member. With modestly reduced levels of ing4, brd1, and jade3 individually, we see no effect on HSC specification. However, when combined we see a loss of marker expression, suggesting they genetically interact to regulate HSC specification. Using ChIP-seq for ING4 in human CD34+ cells, we show that ING4 is bound to many regulators of blood development including c-myb, lmo2, runx1 and ikaros. Ing4 is also bound to several NF-kB target genes including il-1b, il-6, il-8, and il-20, as well as a subset of ifna associated genes. Zebrafish lacking ing4 expression show an increase in NF-kB target gene expression, suggesting that loss of Ing4 results in an overabundance of NF-kB target gene signaling. Mice lacking Ing4 also have altered hematopoiesis. These mice exhibit a differentiation block in short-term HSCs (ST-HSCs) and increased NF-kB target gene expression in HSCs and progenitor cells. Loss of Ing4 has a cell autonomous effect on HSCs where long-term HSCs are unable to repopulate recipient animals in competitive transplantation assays. Surprisingly, Ing4 null multipotent progenitors (MPPs) cells outperform their wildtype counterparts in competitive transplantation, suggesting that MPPs that overcome the block at the ST-HSC stage are superior in repopulation of the niche. Concomitant loss of Ing4 and NF-kB target genes by morpholino can rescue HSC specification in zebrafish. These results suggest a novel mechanism for HSC regulation by Ing4 and the Hbo1 complex through regulation of inflammatory signaling.