Fig. 4: Lactate and α-KG are involved in liver TIC self-renewal.
A CD133+ TIC ratios after 2 days’ treatment with the indicated metabolites were detected via FACS. DM-αKG, cell permeable α-KG. n = 4 independent experiments. B Typical images and sphere diameters in lactate and DM-αKG treated sample #2 cells. n = 30 spheres from three independent experiments were measured. D3, day 3. Scale bars, 100 μm. C Sphere formation of sample #2 cells supplemented with indicated levels of lactate and DM-αKG. n = 4 independent experiments. Scale bars, 500 μm. D Quantitative real-time PCR detection for the indicated gene expression in lactate and DM-αKG treated primary sample #2 cells. n = 3 independent experiments. E Xenograft tumors were established in BALB/c nude mice and treated with FX-11 or DM-αKG after tumors reached about 250mm3, and tumor volume was detected every 3 days. n = 6 6-week-old male BALB/c nude mice per group. F CD133 immunohistochemistry in Vehicle (Veh), lactic acid production inhibitor FX-11 and DM-αKG treated tumors. n = 6 tumors per group, and for each tumor, n = 10 images were detected. Scale bars, 50 μm. G 10, 1 × 102, 1 × 103, 1 × 104 and 1 × 105 Huh7 sphere cells were injected into BALB/c nude mice for tumor initiation assay. n = 6 6-week-old male BALB/c nude mice per group. TIC ratios and P-values calculated via ELDA were shown in the right panels. H 1 × 106 luciferase labeled primary #2 cells were used for in vivo propagation, and mice were administered with FX-11 (2 mg/kg) or DM-αKG (500 mg/kg). Typical liver images and calculated results were shown. n = 6 mice per group. For A, C, n = 4 independent experiments. In all panels, data are shown as mean + s.d. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001, by two-tailed Student’s T-test. Source data are provided as a Source Data file.
