Extended Data Figure 5: Conditional deletion of Dnmt3a in activated CD8 T cells inhibits effector-associated de novo DNA methylation but does not impair maintenance methylation.
From: Effector CD8 T cells dedifferentiate into long-lived memory cells
a, Cre recombinase expression is driven by the Gzmb promoter to initiate recombination of Dnmt3a exon 19 following T cell activation. b, Representative FACS analysis of virus-specific CD8 T cells sorted 8 days after acute viral infection of wild-type and Dnmt3a cKO mice. c, Recombination of genomic DNA from FACS-purified Dnmt3a cKO virus-specific CD8 T cells was assessed by PCR using primers that anneal to DNA outside the floxed target region. The larger PCR amplicon corresponds to the intact locus and the smaller PCR product is the amplicon of the recombined locus. d, Representative and graphical summary of Sell promoter methylation in wild-type and Dnmt3a cKO cells. Mean and s.d. were calculated from bisulfite sequencing analysis of six individually sorted populations. e, Diagram of Sell promoter CpG location proximal and distal to the TSS. f, Representative DNA methylation analysis of CpG sites distal to the Sell promoter regions in day 8 wild-type and Dnmt3a cKO antigen-specific effector CD8 T cells. Graphical summary of the average Sell distal CpG methylation in wild-type and Dnmt3a cKO cells calculated from bisulfite sequencing analysis of four individually sorted populations.
