Extended Data Fig. 7: PE regulates CDP–ethanolamine pathway-mediated TFH responses.
From: Metabolic control of TFH cells and humoral immunity by phosphatidylethanolamine
a, Wild-type and Pcyt2-deficient TFH cells were sorted from LCMV-infected mice and incubated with [3H]-Etn (2 μCi ml−1) for 3 h. Lipids were extracted and [3H]-Etn incorporated into TFH cells was assessed with a scintillation counter (n = 4 samples). b–e, Lipid add-back–rescue assay. The sgNTC and sgPcyt2-transduced SMARTA cells were supplemented with the indicated lipids, followed by transfer of these cells to C57BL/6 mice and LCMV infection. b, Diagram showing the time points of lipid treatment. PE was supplemented at days −5, −3 and −1 before adoptive transfer, and LCMV-infected mice were analysed at day 3 after infection (to capture the effects induced by in vitro PE supplementation). c, Lipidomic analysis showing the PE content in the indicated groups (left), and Venn diagram showing the rescued PE molecules by diacyl-type and ether-type PE supplementation (right) at day 3 after infection. d, Analysis and quantification of the proportion of PE+ cells in donor-derived TFH cells (PSGL-1−Ly6C−) from the spleen (n = 5 mice). e, Analysis and quantification of the proportion of donor-derived CXCR5+SLAM− TFH cells from the spleen (n = 5 mice). f, GSEA plot for KEGG signature of ABC transporters in TFH versus TH1 cells (from a public dataset GSE74854 (ref. 37)). g, Heat map showing the top 22 leading-edge genes from the enrichment plot in f. Data are representative of two (a, c) or at least three (d, e) independent experiments. Data are mean ± s.e.m. P values are determined by one-way ANOVA (d, e) or two-tailed unpaired Student’s t-test (a). *P < 0.05, **P < 0.01 and ***P < 0.001. Numbers in gates indicate percentage of cells.
