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. 2022 Aug 15;18(14):5260–5275. doi: 10.7150/ijbs.73890

Figure 3.

Figure 3

Celastrol shifted macrophage metabolism from glycolysis to mitochondrial respiration against LPS stimulation. (A) Assay for extracellular acidification rate (ECAR). (B) Quantification for the basic and maximum capacity of ECAR. (C) Assay for oxygen consumption rate (OCR). (D) Quantification for the basic and maximum capacity of OCR. Briefly, the cells were incubated with celastrol in the presence of LPS for 12 h. Assays for ECAR and OCR were performed with RAW264.7 cells according to Agilent Seahorse analysis guidelines. (E) Assays for glucose consumption and lactate production. The contents of glucose and lactate were measured using the commercial kits. (F) Inhibition of glycolytic proteins by celastrol. After treated celastrol and LPS as stated in “Methods”, RAW264.7 cells were lysed and examined by WB analysis. (G) Quantitative analysis results of glycolytic proteins (n=3). (H) Inhibition of glycolysis-regulating proteins (e.g., p-AKT, p-mTOR and HIF-1α) by celastrol. (I) Quantitative analysis results of glycolysis-regulating proteins (n=3). * p < 0.05, ** p < 0.01, ***p < 0.001 (LPS vs others).