PD-L1 Expression in Human Breast Cancer Stem Cells Is Epigenetically Regulated through Posttranslational Histone Modifications

Figure 2

Analysis of CpG methylation status and expression of methylation/demethylation enzymes in MCF-7 and BT-549 cell lines and tumorspheres. Representative plots of PD-L1 promoter CpG methylation status analyzed by bisulfite sequencing of the genomic DNA isolated from MCF-7 (a) and BT-549 (b) cell lines and tumorspheres. Methylation status of individual CpG motif is shown by white (demethylation) or gray (methylation) colors. Bar plots show the methylation percentage of PD-L1 (c). Bar plots show the relative expression of DNMT3a, DNMT3b, TET1, TET2, and TET3 in MCF-7 (d) and BT-549 (e) cell lines and tumorspheres. All data were normalized to β-actin. Bar plots show the relative enrichment of 5-mC and 5-hmC in the PD-L1 promoter of MCF-7 (upper) and BT-549 (lower) cell lines and tumorspheres (f). Data are representative of two independent experiments.

(a)

(b)

(c)

(d)

(e)

(f)