Fig. 5

Erythroid differentiation is associated with high-level Setd8 expression. a Levels of SETD8, GATA1, and H4K20me1 during erythroid differentiation of CD34+ HSPCs. Cultures were serially monitored by flow cytometric analyses for Glycophorin A (GYPA) expression and the levels of SETD8, GATA1, and H4K20me1 determined by western blotting. Equivalent protein content was loaded in each lane and total H4 was used as a loading control. b Colony forming assays following treatment of CD34+ HSPCs with the Setd8 inhibitor UNC0379. n = 3 replicates. Error bars represent standard error of the mean (SEM). Significance determined via students T test. c Deletion of the Setd8 + 4.4 enhancer results in decreased mRNA (left) and protein expression (right), as well as reduced levels of H4K20me1. *Indicates suspected non-specific cross-reacting band. Three independent enhancer deletion clones were analyzed. Error bars represent SEM. Significance determined via students T test. d Colony forming assays following genome editing for the SETD8 erythroid enhancer in CD34+ HSPCs. N = 3 replicates. Error bars represent SEM. Significance determined via students T test