Journal: Cancer Immunology, Immunotherapy : CII
Article Title: Metformin drives HIF-1α–mediated dual metabolic reprogramming to enhance γδ T cell therapy in triple-negative breast cancer
doi: 10.1007/s00262-026-04351-w
Figure Lengend Snippet: Metformin exerts distinct effects on OXPHOS and glycolysis in γδ T cells versus TNBC cells through HIF-1α. A , B OCR profiles of TNBC cells and γδ T cells following 24-h metformin treatment, measured by Seahorse XF metabolic flux analysis. OCR curves were assessed under basal conditions and after adding the indicated mitochondrial inhibitors: oligomycin, FCCP, and rotenone/antimycin. C , D ECAR of TNBC cells and γδ T cells were quantified after 24 h of metformin exposure, with measurements taken during basal states and following sequential administration of glucose, oligomycin, and 2-DG. E Representative immunoblots showing AMPK signaling in TNBC and γδ T cells after 4 h exposure to increasing metformin doses. β-actin was employed for normalization. F , G OCR and ECAR profiles of γδ T cells following 24-h metformin and PX-478 treatment, measured by Seahorse XF metabolic flux analysis. All experimental procedures included a minimum of three independent biological replicates. Data represent mean ± SEM; statistical significance determined by two-tailed Student’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001, ns, not significant. PX-478: HIF-1α Inhibitor
Article Snippet: The membrane was blocked with 5% non-fat milk in TBST (JXF003, Jingxin Bio) for 1 h at room temperature and subsequently incubated overnight at 4 °C with primary antibodies targeting phospho-AMPKα (2535 T, CST), AMPKα (2793S, CST), HIF-1α (2178S, CST), and β-actin (4970S, CST).
Techniques: Western Blot, Two Tailed Test