Fig. 3

CKLF1-induced glycolysis and inflammation are dependent on activation of the AMPK–mTOR–HIF-1α pathway. A Representative image of the AMPK–mTOR–HIF-1α pathway. Microglia were treated with LPS + IFN-γ and various doses of C27 for 24 h, and immunoblot analysis of mTOR, p-mTOR, HIF-1α, AMPK, p-AMPK, TREM2, IL-6 and β-actin was performed (n = 3 per group). B Quantification of the gray values in A (n = 3 per group). p-AMPK/AMPK: F (4, 10) = 36.37; p-mTOR/mTOR: F (4, 10) = 12.76; HIF-1α/β-actin: F (4, 10) = 124.8; TREM2/β-actin: F (4, 10) = 9.472; IL-6/β-actin: F (4, 10) = 10.59. C Fluorescent signals of TREM2 induced by C27 (n = 3 per group). Scale bar = 100 μm. D Relative mRNA expression of IL-1β, IL-6 and TNF-α in microglia treated with C27 and rapamycin or metformin for 24 h (n = 6 per group). IL-1β: Vehicle: F (5, 5) = 41.99, MET: F (5, 5) = 17.03; IL-6: Vehicle: F (5, 5) = 8.296, MET: F (5, 5) = 1.189; TNF-α: Vehicle: F (5, 5) = 19.24, MET: F (5, 5) = 6.875. IL-1β: Vehicle: F (5, 5) = 9.089, RAP: F (5, 5) = 1.266; IL-6: Vehicle: F (5, 5) = 24.6, RAP: F (5, 5) = 1.601; TNF-α: Vehicle: F (5, 5) = 9.809, RAP: F (5, 5) = 2.741. E Schematic diagram of the effect of rapamycin or metformin on the AMPK–mTOR–HIF-1α pathway. The data are presented as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 vs. control group