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. 2025 Jun 24;152(8):537–554. doi: 10.1161/CIRCULATIONAHA.125.073691

Figure 4.

Figure 4.

One-carbon metabolism is crucial for CD34-lineage CD4+T cell differentiation into T follicular helper cells. A, Monocle pseudotime trajectory of all CD34-lineage T cells. Cells are labeled by pseudotime (left) and cell subtypes (right). B, Heatmap of the significantly changed genes (P<0.01) discovered by the BEAM function from Monocle in branchpoint 1. C, RNA velocity analysis distinguished velocity vectors across the pseudotime axis. D, Monocle pseudotime trajectory of CD34-lineage CD4−CD8− T, CD4+CD8+ T, CD4+ T, and CD8+ T cells of a mouse 2-week remodeled artery after allograft. Cells are labeled by pseudotime (left) and cell subtypes (right). E, monocle pseudotime trajectory of CD34-lineage CD4−CD8− T, CD4+CD8+ T, CD4+ T, and CD8+ T cells of a mouse 4-week remodeled artery after allograft. Cells are labeled by pseudotime (left) and cell subtypes (right). F, Monocle pseudotime trajectory of CD34-lineage CD4+ T, regulatory T, T follicular helper, and T helper 17 cells of a mouse 2-week remodeled artery after allograft. Cells are labeled by pseudotime (left) and cell subtypes (right). G, Monocle pseudotime trajectory of CD34-lineage CD4+ T, regulatory T, T follicular helper, and T helper 17 cells of a mouse 4-week remodeled artery after allograft. Cells are labeled by pseudotime (left) and cell subtypes (right). H, Volcano plots show the genes with different expression between CD4+ T and CD8+ T cells of a mouse 2-week (left) and 4-week (right) remodeled artery after allograft. Differentially expressed genes (DEGs) are highlighted in red or blue and were determined by Wilcoxon rank-sum test. I, Volcano plots showing the genes with different expression between CD4+ T and T follicular helper cells. DEGs are highlighted in red or blue and were determined by Wilcoxon rank-sum test. J, Heatmap summary of the expression of select metabolic pathway activation–related genes (rows) in the indicated cell types (columns), as determined by single-cell RNA sequencing. K, Heatmap summary of the expression of select metabolic pathway activation–related genes (rows) in the indicated T cell subtypes (columns), as determined by single-cell RNA sequencing. L, Schematic illustrating the conclusion that several metabolic pathways are activated during differentiation of CD34-lineage CD4+ T cells into T follicular helper cells. Th17 indicates T helper 17; Tm, memory T; Tn, naïve T; and Treg, regulatory T.