DMS validation and distribution of DMSs. (A) Correlation plot comparing the β-values for the 10 156 DMSs in PMNs with published 450K data from peripheral neutrophils.35 The plot shows the average of the 3 PMN replicates from this study vs the average of the 6 neutrophil replicates. Lower density, red; higher density, blue. The Pearson correlation coefficient is given above the plot. (B) Correlation plot comparing the average β-values of 16 DMSs in CMP, GMP, and PMC, with pyrosequencing results (average of biological triplicates). The DMSs were located in the MPO, PRTN3, AZU1, ELANE, GFI1, TBCD, and HDAC4 genes. One additional 450K CpG (probe cg07805029) in GFI1 was also included in pyro analysis but not in the list of DMSs (P = .07). The Pearson correlation coefficient is given above the plot. (C) Bar chart showing the methylation levels of the DMSs plotted in B (including the additional probe cg07805029) in GMP (white) and PMC (gray) measured by bisulfite pyrosequencing. The bars show the average of biological triplicates and error bars display standard deviation. Red dots mark the corresponding average β-value from 450K analysis. The 450K probe ID is stated on the x-axis. (D) The bar chart displays the relative distribution of CpGs over CpG islands, shores, shelves, and open sea. Blue bars represent CpGs present on the 450K methylation array and green bars represent DMSs. The numbers of CpGs are given in the adjacent table. Total number of cytosines on array after filtering for detection P < .01 was 477 112. Total number of differentially methylated CpGs was 10 156. P values were calculated with the χ2 test.
