Fig. 1

CKLF1 activates microglia and induces metabolic reprogramming to aerobic glycolysis. A Time course of IL-6 protein expression in response to C27 stimulation (n = 3 per group). F (9, 20) = 63.38. B Microglial phagocytosis was activated by C27 exposure. Microglial phagocytosis was tested using imaging and flow cytometry in the control group, positive control group (LPS + IFN-γ) and low-dose C27-L (500 nM), medium-dose C27-M (1000 nM), high-dose C27-H (2000 nM) administration groups (C27-L, C27-M, C27-H). The amount of zymosan that was phagocytosed by microglia was determined by fluorescence imaging of single cells. Scale bar: 7 μm. The proportion of zymosan phagocytosed by a single cell in each group (n = 3 per group). 1 bead: F (4, 10) = 8.54; 2 beads: F (4, 10) = 11.59; 3 beads: F (4, 10) = 5.078; 4 beads: F (4, 10) = 9.009. C, D Microglia glycolysis was enhanced by C27. The cell energy metabolism analyzer can detect real-time changes in extracellular acidification and quantitatively determine basic glycolytic capacity, maximum glycolytic capacity and reserve glycolytic capacity to measure the ECAR (n = 4 per group). Basal glycolysis: F (4, 15) = 51.77; Max glycolysis: F (4, 15) = 68.4; Reversed glycolysis: F (4, 15) = 3.952. E The cellular energy metabolism analyzer detects the real-time changes in the oxygen consumption rate of microglia (n = 4 per group). F Quantitative statistics of the basal respiration, proton leak and maximum oxygen consumption rate (n = 4 per group). Basal respiration: F (4, 15) = 9.005; Proton leak: F (4, 15) = 1.674; Maximal respiration: F (4, 15) = 14.82. G Relative mRNA expression of glycolytic genes stimulated by C27 (n = 5–6 per group). GLUT1: F (4, 25) = 2.787; HK: F (4, 25) = 8.762; PFKFB3: F (4, 20) = 7.334; PKM2: F (4, 25) = 3.341. H Schematic diagram of the metabolic reprogramming process induced by C27. The data are presented as the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 vs. control group