Quantitative RT-PCR of mRNA isolated from cultured human primary erythroid cells. Q-PCR was performed using iQ SYBR Green Supermix (BioRad). Relative expression was normalized to geometric mean of unvarying, ubiquitously genes, ornithine decarboxylase antizyme 1 (OAZ1), hypoxanthine phosphoribosyltransferase 1(HPRT1), ubiquitin-fold modifier 1 (UFM1), TATA box binding protein (TBP), pumilio homolog 1 (PUM1), and ribosomal protein S13 (RPS13) as controls. The changes in specific mRNA levels were calculated using the ΔΔC_T method (where C_T is threshold cycle), with results presented as means ± SEM. Results were normalized to the gene with the highest expression level in each group. Triplicate analyses were performed for each target gene.