Expression data from knockdown of ZXDC1/2 in PMA-treated U937

ZXDC1 augments the expression of various markers of monocyte/macrophage differentiation when over-expressed in the U937 cell line treated with the phorbol ester PMA. Likewise, knockdown of ZXDC1 restricts the induced expression of these markers. We sought to identify specfic gene targets of ZXDC1 during the process of monocyte/macrophage differentiation in U937 by performing gene expression profiling in cells exhibiting reduced expression of ZXDC1 compared to controls. We used microarray to identify genes regulated by ZXDC during PMA-induced differentiation of U937 monoblasts towards a monocyte/macrophage phenotype An inducible shRNAmir (Tet-On) cell line was established in the U937 background that displayed significant, reproducible, and doxycycline inducible knockdown of ZXDC1. Using this cell line, we created four experimental groups to identify gene targets of ZXDC1 during PMA-induced differention towards a monocyte/macrophage phenotype: Vehicle alone; Vehicle+Doxycycline; PMA alone; PMA+Doxycycline. Each independent group was used for RNA extraction and hybridization on Affymetrix microarray chips.