Effect of HES6 knockdown on gene expression in vitro differentiated megakaryocytes, erythroblasts and precursor cells, cultured from CB CD34+ HSPCs.

Cord blood CD34+ hematopoietic precursors (3 donors) were control or HES6 shRNA (HES6 knockdown) transduced and cultured for four days in media containing SCF, TPO and EPO to drive differentiation towards megakaryocytes and erythroid cells. After four days four subpopulations were sorted for RNA isolation to research the effect of HES6 knockdown on gene expression. We sorted megakaryocytes (CD41+) and early (CD71+ CD235-) and late (CD71+ CD235+) erythroblasts (CD41-) and CD34- precursor cells (CD41- CD71- CD235- CD34-). Overall design: Gene expression profiling by high throughput sequencing of 4 different cell types (megakaryocytes, CD34- precursors, early and late erythroblasts) for 3 replicates with control or HES6 knockdown (2 conditions: shRNA1 and shRNA2, both targeting human HES6).