Journal: Nature

Article Title: Activated ATF6α is a hepatic tumour driver restricting immunosurveillance

doi: 10.1038/s41586-025-10036-8

Figure Lengend Snippet: ( a-b ) Metabolic flux analysis ( a ) for extracellular acidification rates (ECAR) of FL83B WT (n = 3) or FL83B TG (n = 3) hepatocytes, with quantification ( b ) for glycolysis and glycolytic capacity. ( c ) Glucose consumption in FL83B WT or FL83B TG hepatocytes cultured for 48 h (n = 6/group). ( d ) qRT-PCR analysis of glucose-related metabolic genes in FL83B WT or FL83B TG cells (n = 6/group). Bold labelling depicts significant genes. ( e ) NMR spectroscopy-based metabolomics analysis of lactate concentration in FL83B cells with ATF6α expression (FL83B TG ) or deletion (FL83B KO ), normalized to respective controls (n = 6/group). ( f ) NMR metabolomics PLS − DA with coloured 95% confidence intervals illustrating the clear group separation in metabolic profile between FL83B WT and FL83B TG cells (n = 6/group). ( g ) qRT-PCR analysis of primary hepatocytes harvested from livers of 3-month-old TG Alb-cre− (n = 4) and TG Alb-cre+ (n = 6) mice. Bold labelling depicts significant genes. ( h ) qRT-PCR analysis of primary hepatocytes harvested from livers of CD-HFD-fed Atf6 fl/fl (n = 6) and Atf6 ΔHep (n = 6) mice. Bold labelling depicts significant genes. ( i ) Scheme of HCC cancer cell line (HLE) stably transfected with control vector (HLE WT ) or nATF6α-expressing (HLE TG ) plasmid and co-cultured with or without cytotoxic MART-1 T-cells to assess immune attack by a real-time cell analyzer (xCELLigence). ( j ) HLE WT or HLE TG cancer cell growth over time with quantification on the right panel (n = 3/group). ( k ) Killing assay and quantification for HLE WT or HLE TG cancer cell growth in co-culture conditions with cytotoxic MART-1 T-cells (n = 3/group). ( l ) Scheme of Colo800 tumour cells stably transfected with control (Colo800 WT ) or nATF6α-expressing (Colo800 TG ) plasmid and co-cultured with cytotoxic MART-1-specific T-cells to assess immune attack in a real-time cell analyzer (xCELLigence). ( m ) Killing assay of Colo800 WT or Colo800 TG tumour cells in co-culture conditions with cytotoxic MART-1-specific T-cells (n = 3/group) in the presence or absence of the lactate dehydrogenase inhibitor galloflavin. ( n ) Killing assay for Colo800 WT or Colo800 TG tumour cell growth in co-culture conditions with cytotoxic MART-1 T-cells (n = 4/group) in the presence or absence of the lactate efflux inhibitor AZD3965. ( o ) Glucose consumption of Colo800 WT and Colo800 TG cells cultured for 48 h (n = 6/group). ( p ) qRT-PCR analysis of indicated mRNAs from Colo800 WT and Colo800 TG cells (n = 6/group). Bold labelling depicts significant genes. ( q ) NMR metabolomics PLS − DA with coloured 95% confidence intervals illustrating the clear group separation in metabolic profile between Colo800 WT (n = 5) and Colo800 TG (n = 6) cells. ( r ) Top 15 metabolites and their VIP score based on the PLS − DA regression model. Scatter dot plot data are presented as mean values ± SEM. Line graph data are presented as mean values ± SEM (10a) or mean values ± SD (10j-k, m-n). Data in 10b-d,g,h,o,p were analysed by two-tailed unpaired t -test or Mann-Whitney test based on data normality distribution. Data in 10e were analysed by one-way ANOVA. Data in 10j,k were calculated for the area under the curve and analysed by two-tailed Student’s t -test. Data in 10m,n were calculated for the area under the curve and analysed by one-way ANOVA. In vitro co-culture schematics were created in BioRender. Heikenwälder, M. (2026) https://BioRender.com/lgjnsy9 .

Article Snippet: Clear supernatant was filled into 1.7 mm Bruker SampleJet-compatible NMR spectroscopy tubes.

Techniques: Cell Culture, Quantitative RT-PCR, Structural Proteomics, Concentration Assay, Expressing, Stable Transfection, Transfection, Control, Plasmid Preparation, Co-Culture Assay, Two Tailed Test, MANN-WHITNEY, In Vitro