Extended Data Fig. 6: Signalling pathways of TCF1 and LEF1 on mice and human B-1 cells.

(a) Expression-expression plot of transcriptomes of B-1a cells isolated from TCF1^WTLEF1^WT and TCF1^ΔLEF1^Δ mice. Superimposed B-1a signature genes from published RNA-seq data which over- or under-represented in peritoneal B-1a versus B-1b cells³⁴. (b) Expression heatmap of B-1a signature genes, with the colour representing normalized RPM for each gene across different samples. (c) Dot plot presentation of Hallmark gene sets according to GSEA for differentially expressed genes in TCF1^WTLEF1^WT and TCF1^ΔLEF1^Δ B-1a cells. (d) Cnet plot presentation of enriched Hallmark pathways (P < 0.05, FDR < 0.05) upon GSEA. Size = the number of differentially expressed gene which belongs to the enriched Hallmark pathway. Fold change difference between TCF1^WTLEF1^WT and TCF1^ΔLEF1^Δ B-1a cells is shown. (e) Frequency of TCF1 and LEF1 expressed cells in human B-1 clusters²³. (f) Dot plot representation of Tcf7 and Lef1 mRNA expression in different subsets of human prenatal tissues. Panels e,f were adapted from ref. ²³, AAAS. (g) Flow cytometry plots and quantification of B-1-enriched cells from PBMCs from 4 SCID patients and 3 HD. (h) ChIP-seq analysis of TCF1 binding in CD4⁺CD8⁺ thymocytes⁴² and ATAC-seq analysis at Cd5, Myc and Ets1 on B-1a cells from Vh12/Vk4 transgenic mice¹⁷. Panel h was adapted from refs. ^17,42, Springer Nature Ltd. The significant hallmark gene sets in MSigDB (adjustment p-value < 0.05) for each contrast based on the hypergeometric testing using cluster profiler package (c).

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