Skip to main content
. Author manuscript; available in PMC: 2016 Apr 1.
Published in final edited form as: Transfusion. 2014 Nov 2;55(4):864–874. doi: 10.1111/trf.12904

Figure 2. Global methylation and gene transcript levels in primary uncultured MPB CD34+ cells and after treatment with 5azaD/TSA.

Figure 2

Long interspersed nucleotide element-1 (LINE-1 assays) assays were performed using genomic DNA to compare global methylation of four CpG sites between purified CD34+ cells derived from unmanipulated primary cord blood, bone marrow and mobilized peripheral blood (MPB) samples and at 72 hours after treatment of MPB cells with or without 5azaD/TSA.

A) Global methylation levels were similar between unmanipulated, primary CD34+ cells from cord blood, bone marrow, and MPB samples. *The data on cord blood in part was previously published (Ref #37, reprinted with permission) and used here for comparison purposes. B) Global methylation levels decreased significantly after sequential treatment with 5azaD/TSA compared to cells cultured in cytokine-only conditions (53.7 ± 4.5% vs. 74.0 ± 7.9%, respectively; P =0.004). C-F) Relative mRNA levels in MPB cells cultured in 5azaD/TSA and Cytokine A show higher transcript levels of HOXB4, EZH2, and GATA2 in 5azaD/TSA treated cells and lower transcript levels of PU.1 compared to Cytokine A-only expanded cells. Gene transcript levels were measured using the whole culture product derived from CD34+ cells after 9 days of culture. Measurements were obtained in triplicate and a negative control (lacking the cDNA template) was included for each assay (n = 3).