Modulation of histone modifications by specific inhibitors. Naive cord blood CD4 T cells were stimulated with anti-CD3/28 in the presence of the indicated cytokines and the presence or absence of increasing concentrations of DZNep or curcumin. (A) Expression of PU.1, IRF4, T-bet, GATA-3, and RORC mRNA were normalized for the expression of cyclophilin. Results are shown in relation to mRNA expression in untreated controls. (B) Expression of IL-9 mRNA was normalized for the expression of cyclophilin. (C) In addition to DZNep treatment, cells were transfected with control, scrambled (scr), or PU.1-specific siRNAs. PU.1 protein expression was assessed by flow cytometry 5 days after anti-CD3/CD28 stimulation. PU.1 and IL-9 mRNA expression were assessed by PCR. mRNA levels were normalized to cyclophilin. The mean ± SEM of at least 3 independent experiments with cells from different donors is shown. *P < .05; **P < .01; ***P < .001; n.s. indicates not significant by Student t test.