Fig. 5: VLCFA reduction mediates INSIG1 instability and SREBP2 activation. | Nature Metabolism

Fig. 5: VLCFA reduction mediates INSIG1 instability and SREBP2 activation.

From: A functional single-cell metabolic survey identifies Elovl1 as a target to enhance CD8+ T cell fitness in solid tumours

Fig. 5

a, Bulk-RNA sequencing analysis of biological process significantly upregulated in vitro in sgElovl1 (n = 2) compared with sgNT (n = 2) OT-I T cells on day 7 from activation (day of ACT). b, Representative western blot (left) and quantification (right) of INSIG1 in sgNT, sgElovl1 and sgElovl1 supplemented with 20 µM C24 OT-I T cells in vitro (n = 4) on day 5 from activation. c, Representative flow cytometry plots and quantification (correlation SREBP2/ 7-AAD) of SREBP2 nuclear translocation in sgNT or sgElovl1 OT-I T cells in vitro (n = 3) on day 5 from activation performed by BD FACSDiscover S8 Cell Sorter. d, Srebp2 and Hmgcr expression assessed by RT–qPCR in vitro in sgNT or sgElovl1 OT-I T cells (n = 5) on day 5 from activation. e,f, in vitro flow cytometry quantification of LDLR (e) and FILIPIN III (f) in sgNT and sgElovl1 OT-I T cells ± lignoceric acid 20 µM (C24), 2 µM Lovastatin or 2 µM R0 48-8071 (n = 4) on day 5 from activation for 24 h. g, GSEA of cholesterol biosynthesis and uptake gene expression evaluated via bulk-RNA sequencing of sgNT (n = 3) or sgElovl1 (n = 3) OT-I T cells sorted from PDAC primary tumour of mice treated with anti-PD-1. h, Tumour weight on day14 post-KPC_OVA injection, of mice that received sgNT (n = 12), sgElovl1 (n = 11) or sgElovl1 pretreated with 2 µM Lovastatin (n = 5) OT-I T cells and treated with anti-PD-1. i, Tumour weight on day14 post-KPC_OVA injection, of mice that received sgNT (n = 8), sgElovl1 (n = 7) or sgElovl1 pretreated with 20 µM C24 (n = 10) OT-I T cells and treated with anti-PD-1 (two independent experiments). j,k, Flow cytometry quantification of SLAMF66+TIM3− (j, Tpex) and IFNγ+TNF+IL-2+ (k, T polyfunctional) in sgNT, sgElovl1 or sgElovl1 pretreated with 20 µM C24 OT-I T cells infiltrating the primary tumour of KPC_OVA-bearing mice treated with anti-PD-1 (sgNT n = 10, sgElovl1 n = 9, sgElovl1 + C24 n = 9, two independent experiments). Data are presented as mean ± s.e.m. Statistical significance was assessed by paired (c,d) two-tailed Student’s t-test or one-way ANOVA (b,f,h–k). NES, normalized enrichment score.

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