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. Author manuscript; available in PMC: 2019 Jun 10.
Published in final edited form as: Nat Immunol. 2018 Dec 10;20(1):73–85. doi: 10.1038/s41590-018-0274-0

Fig. 6. c-Maf is essential for Tγδ17 programming.

Fig. 6.

(a) Differential mRNA expression in Maf+/+Il7rCre (WT) and Maffl/flIl7rCre (KO) E17–18 CD25−CD27− γδ thymocytes displayed as a volcano plot of log2 fold change vs. the –log10(p-value) for each gene. Genes considered significant (FDR < 0.05) are in orange, while select Tγδ17-associated genes are in blue. p-value capped at 10−40. (b) RNA expression values for known Tγδ17 transcriptional regulators from (a) RPKM, reads per kilobase million. ♦, significant differential expression at FDR < 0.05. (c) Differential ATAC-seq analysis in Maf WT and KO E17–18 CD24+CD45RBloTCRγδ+CD3ε+ cells. Regions having significantly differential accessibility (FDR < 0.05) are in orange. Significant regions within 10 kb up- or down-stream of a differentially-expressed gene (Maf KO) are further highlighted in blue, with select type-17-associated regions annotated with gene symbols, p-value capped at 10−50. Proportion DA of total significant ATAC peaks indicated for each half (top). Results of de novo motif enrichment analysis comparing differential ATAC peaks (FDR<0.05) against non-differential, with the two most-enriched motifs shown (bottom). (d) ATAC-seq tracks at select loci displayed using IGV for Maf WT and KO CD24+CD45RBlo γδ thymocytes. ΔATAC* track and shading denotes DA regions (FDR<0.05). MARE and RORE indicated in blue, MARE with human conservation in red, location of qPCR amplicons in green. (e) Comparison of differential expression log2 fold-change by RNA-seq in Maf KO and Rorc(t) KO E17–18 CD27−CD25− γδ thymocytes. Genes differentially expressed (FDR < 0.05) in Maf KO (blue), Rorc(t) KO (black), or both (orange) are highlighted, with select type 17 program genes annotated. (f) c-Maf and RORγt ChIP-qPCR analysis of the Il17a locus, and (g) c-Maf ChIP-qPCR of the Blk and Tcf7 loci for WT and KO E18 CD45RBlo γδ T cells Number of biological replicates per group for (a-d) n=2, (e) n=4, (f, g) n=3. Mean ± SEM used. * p<0.05, ** p<0.01, *** p<0.001, two-way ANOVA with Fisher’s LSD post-test (f left, g).