Combination of EHMT1/2 pharmacologic inhibition or knockdown with entinostat or decitabine shows additive effects on γ-globin gene induction in primary adult human erythroid cells. (A) Histograms showing the relative percentage of γ-globin genes expression according to the concentration of drug(s). qRT-PCR analysis was performed at day 14 of erythroid differentiation, after treatment with UNC0638 only, entinostat (MS-275) only, or UNC0638 + MS-275 for 7 days (from day 7 to day 14) (mean ± SD, n = 5 biological replicates). ***P < .001. (B) Histograms showing the relative percentage of γ-globin genes expression according to the concentration of drug(s). qRT-PCR analysis was performed at day 14 of erythroid differentiation, after treatment with UNC0638 only (from day 7 to day 14), decitabine (DAC) only (from day 11 to day 14), or UNC0638 + DAC (from day 7 to day 14 for UNC0638 and from day 11 to day 14 for DAC) (mean ± SD, n = 5 biological replicates). **P < .01; ***P < .001. (C) Histograms showing the relative percentage of γ-globin genes expression in knockdown cells, in the presence of 0.5 μM MS-275 or the vehicle control for 3 days (from day 11 to day 14). qRT-PCR analysis was performed at day 14 of erythroid differentiation (mean ± SD, n = 4-6 biological replicates). One-way ANOVA, P < .001. (D) Histograms showing the relative percentage of γ-globin genes expression in knockdown cells, in the presence of 0.5 μM DAC or the vehicle control for 3 days (from day 11 to day 14). qRT-PCR analysis was performed at day 14 of erythroid differentiation (mean ± SD, n = 4-6 biological replicates). One-way ANOVA, P < .001.