Transdifferentiation of B cells to macrophages occurs without DNA methylation changes in key genes. (A) Selected examples of bisulfite genomic sequencing at the promoter CpG islands and CpG island shores of two macrophage-specific genes in parental HAFTL B cells, RAW macrophages, and C/EBPαER-expressing B C10 cells (derived from HAFTL) at 0 h (after C/EBPα induction) and reprogrammed into functional macrophages (48 and 120 h). Percentage of DNA methylation is represented by red bars and CpG site number included in the analysis is indicated at the bottom. The two analyzed regions are CpG island (right panel, black bar) and CpG island shore (left panel, grey bar). (B) Selected examples of bisulfite genomic sequencing at the promoter CpG islands and CpG island shores of two B cell-specific genes in parental HAFTL and RAW cells and C10 cells at 0 h and fully reprogrammed into functional macrophages (48 and 120 h). The two analyzed regions are CpG islands (right panel, black bar) and CpG island shores (left panel, grey bar). (C) Heatmap of DNA methylation data for 13 macrophage-specific genes (top) and 11 B cell-specific genes (bottom) in HAFTL and RAW cells and C10 cells at three times during β-estradiol-induced reprogramming (0, 48 and 120 h). Heatmap of DNA methylation data including the four macrophage-specific genes that are differentially methylated between primary B cells and macrophages are also included at the bottom. The number of CpG sites analyzed for each gene is indicated on the right of the heatmap. Values are in Supplementary Table S2. (D) qRT-PCR kinetics of expression of DNA methyltransferases (Dnmt1, Dnmt3a and Dnmt3b) and activation induced deaminase (Aid). H and R, respectively, designate HAFTL and RAW cells. (E) qPCR analysis of MBD-bound (methylated) fractions obtained from MethylCAP experiments with HAFTL and RAW cells and C10 cells at 0, 48 and 120 h for two macrophage-specific and two B-cell-specific genes.