Figure 6. PRMT5 controls key Th17 development, pathogenicity, and metabolic genes and promotes ECAR/OCR during Th17 polarization.

(A) Volcano plot of significantly induced and repressed genes (FDR < 0.05) in Th17 condition–activated naive T cells from T-PRMT5^Δ/Δ versus PRMT5^fl/fl mice (n = 3 each). (B) Heatmap of cholesterol biosynthesis enzymes differentially expressed in CD4⁺ Th17 cells from A, showing similar data to that obtained in Th0 conditions (Figure 5). (C) Heatmap of Th17 differentiation signature genes, as defined by Ciofani et al. (30) and Yosef et al. (31), significantly differentially expressed in T-PRMT5^Δ/Δ versus PRMT5^fl/fl mouse Th17 cells from A. (D) FC in expression of Th17 pathogenic signature genes, as defined by Lee et al. (32) and Gaublomme et al. (33), observed in T-PRMT5^Δ/Δ versus PRMT5^fl/fl mouse Th17 cells from A. (E) STAT3 phosphorylation Western blot and quantification of p-STAT3 to total STAT3 ratio in T-PRMT5^Δ/Δ versus PRMT5^fl/fl Th17 cells. Three independent experiments with 2 to 3 mice/experiment. (F) Glycolysis, lactate, and TCA pathway genes were downregulated in T-PRMT5^Δ/Δ versus PRMT5^fl/fl mouse Th17 cells from A. Circle indicates the percentage of genes within the pathway reduced in T-PRMT5^Δ/Δ T cells. (G) Extracellular acidification rate (ECAR) in Jurkat T cell lines stably transduced with control or PRMT5 shRNA (70%–75% knockdown). (H) Oxygen consumption rate (OCR) in Jurkat T cell lines stably transduced with control or PRMT5 shRNA (70%–75% knockdown). For G and H, the left panel corresponds to lifetime ECAR/OCR signal for 1 representative experiment, with each data point corresponding to 3 technical replicates and the right panel corresponds to data pooled from 4 (ECAR) or 3 (OCR) independent experiments; Student’s t test, *P < 0.05, ***P < 0.001. Bar graphs display mean ± SD.