Fig. 3: IL-27 reduces mitochondrial mass and impedes oxidative phosphorylation in γδ T17 cells. | Cell Death & Disease

Fig. 3: IL-27 reduces mitochondrial mass and impedes oxidative phosphorylation in γδ T17 cells.

From: IL-27 disturbs lipid metabolism and restrains mitochondrial activity to inhibit γδ T17 cell-mediated skin inflammation

Fig. 3

RNA-sequencing analysis was performed using in vitro differentiated γδ T17 cells from the spleen of Cd2-Cre Il27raflox/flox or Cd2-Cre mice. GSEA analysis of genes in mitochondrion (a) and oxidative phosphorylation (b) were shown. c MitoTracker staining of WT γδ T cells that were differentiated in vitro under γδ T17 condition with or without rmIL-27 (50 ng/ml) (n = 6). d Transmission electron microscopy of WT γδ T cells that were differentiated in vitro under γδ T17 condition with or without rmIL-27 (50 ng/ml). The size and morphology of mitochondria were determined and shown (n = 6). e JC-1 staining of γδ T cells from Cd2-Cre Il27raflox/flox (n = 6) or Cd2-Cre mice (n = 8) that were differentiated in vitro under γδ T17 condition with or without rmIL-27 (50 ng/ml). f Immunoblotting of mitochondrial proteins in WT γδ T cells that were differentiated in vitro under γδ T17 condition with or without rmIL-27 (50 ng/ml) (n = 3). g Oxygen consumption rate (OCR) of WT γδ T cells that were differentiated in vitro under γδ T17 condition with or without rmIL-27 (50 ng/ml) was determined via the Seahorse assay. Cumulative data for the basal and maximal respiration were shown. h Oxygen consumption rate (OCR) of in vitro differentiated γδ T17 cells from Cd2-Cre Il27raflox/flox (n = 5) or Cd2-Cre (n = 6) mice was determined via the Seahorse assay. Cumulative data for the basal and maximal respiration were shown. i Flow cytometry of IL-17 levels in γδ T cells from Cd2-Cre Il27raflox/flox or Cd2-Cre mice that were differentiated in vitro under γδ T17 condition in the presence or absence of Oligomycin A (10 μM) (n = 4). j MitoTracker staining of γδ T cells from Cd2-Cre (n = 6) or Cd2-Cre Il27raflox/flox mice that were differentiated in vitro with (n = 4) or without (n = 6) Forskolin (10 μM). k JC-1 staining of γδ T cells from Cd2-Cre (n = 6) or Cd2-Cre Il27raflox/flox mice that were differentiated in vitro with (n = 6) or without (n = 5) Forskolin (10 μM). Data were presented as mean ± SEM, statistical differences were performed using Two-tailed unpaired student’s t-test (c, d, g, h) or One-way ANOVA (e, i–k). *p < 0.05, **p < 0.01, ***p < 0.001, NS not significant.

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