Validation of the regulatory role of UTX on hematopoietic cell migration. (A) UTX shRNA transduced 32D cells showed a significant reduction in UTX expression compared with scramble shRNA transduced 32D cells. (B) 32D cells containing UTX-specific shRNA demonstrated reduced migration compared with wildtype or scramble shRNA transduced 32D cells. Co-overexpression of UTX CDS rescued the migratory phenotype in 32D cells containing shRNA targeting 3′ UTR of UTX mRNA (shRNA-III) but not in 32D cells expressing UTX CDS specific shRNA (shRNA-I, shRNA-II). Coexpression of GFP CDS did not alter migration. (C) JMJD3 shRNA transduced 32D cells showed a significant reduction in JMJD3 expression compared with scramble shRNA transduced 32D cells. (D) 32D cells containing JMJD3 specific shRNA demonstrated no altered migration compared with wildtype or scramble shRNA transduced 32D cells. (E) UTX shRNA transduced Jurkat cells showed a significant reduction in UTX expression compared with scramble shRNA transduced Jurkat cells. (F) Jurkat cells containing UTX-specific shRNA demonstrated reduced migration compared with wildtype or scramble shRNA transduced 32D cells. (G) UTX shRNA transduced Jurkat cells showed no alteration in global level of tri-methylated histone 3 lysine 27 residue (H3K27me3) relative to histone 3 (H3) level compared with scramble shRNA transduced Jurkat cells. (H) Migrated human primary CD34⁺ hematopoietic stem and progenitor cells (HSPC) of 4 different donors showed significantly higher UTX expression levels than nonmigrated cells. **P < .01