Fig. 5: Rewiring the MUFA metabolism triggers cGAS-STING activation in CD4+ T cells. | Communications Biology

Fig. 5: Rewiring the MUFA metabolism triggers cGAS-STING activation in CD4+ T cells.

From: SCD2-mediated monounsaturated fatty acid metabolism regulates cGAS-STING-dependent type I IFN responses in CD4+ T cells

Fig. 5

a, b Western blot analysis of phospho-TBK1 (pTBK1) and total TBK1 from control, TOFA-treated, sgScd2, and sgFads2 Th1 cells (a). Band intensity was determined by image j and summary of three independent experiments was shown (b). c Western blot analysis of pTBK1 and total TBK1 from control, sgScd2, sgScd2/sgTmem173 DKO and sgScd2/sgMavs DKO of Th1 cells. d Western blot analysis of phospho-STING (pSTING) and total STING from control, sgScd2, sgScd2/sgMb21d1 DKO and sgScd2/sgTmem173 DKO of Th1 cells. e qRT-PCR analyses of the relative expression of ISGs in control, sgScd2, and sgScd2/Mavs DKO of Th1 cells. Relative expression (normalized to Hprt) with SD is shown. f qRT-PCR analyses of the relative expression of ISGs in control, sgScd2, sgScd2/sgTmem173 DKO, and sgScd2/sgMb21d1 DKO of Th1 cells. Relative expression (normalized to Hprt) with SD is shown. g qRT-PCR analyses of the relative expression of ISGs in Th1 cells. 50 μM 2-BP was treated for 72 h. Relative expression (normalized to Hprt) with SD is shown. h, i Western blot analysis of pSTING and total STING was performed. 50 μM 2-BP was treated for 72 h (h). 30 μM OA was treated for 72 h (i). j The amount of cGAMP was measured by ELISA. Data are means ± SD. k The amount of cytosolic DNA was measured by Qubit Fluorometer. Data are means ± SD. l qRT-PCR analyses of the relative expression of genomic DNA and mitochondrial DNA in cytosolic DNA derived from control Th1 or sgScd2 Th1 cells. SD is shown. m qRT-PCR analyses of the relative expression of ISGs in Th1 cells. 100 U/ml IFNβ and 30 μM oleic acid were treated for 72 h. Relative expression (normalized to Hprt) with SD is shown. n, o Western blot analysis of pTBK1, total TBK1, pSTING, STING from Th1 cells was performed. 100 U/ml IFNβ, 30 μM oleic acid, 30 μM palmitic acid or 10 μM stearic acid were treated for 72 h (n). The summary of relative intensity was shown. Band intensity was determined by image (j) and summary of three independent experiments was shown (o). p MLE-15 cells were cultured with culture medium of control, sgScd2, sgScd2/sgMB21d1 DKO, sgScd2/sgTmem173, or sgScd2/sgMavs DKO Th1 cells and infected with x31. Virus titers were determined 72 h p.i. with similar results. More than three independent experiments were performed and showed similar results (a–j and l–p = 3, k = 4). Three technical replicates were performed with quantitative RT-PCR and relative expression (normalized to Hprt or 18S) with SD is shown (e–g and l, m).

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