Research Paper Volume 12, Issue 10 pp 9534—9548

Figure 3. Enhancement of glycolysis reverses the anti-inflammatory effect of DEX on LPS-treated macrophages. (A and B) BMDMs were seeded in Seahorse XFe96 cell culture microplates and treated with 25 ng/ml GM-CSF for 24 h before being treated with 100 ng/ml LPS and 1 μM DEX for 4 h. The real-time ECAR was recorded, and basal glycolysis and glycolytic capacity values were plotted. n = 5; mean ± SEM; * P < 0.05. (C) BMDMs were treated with 25 ng/ml GM-CSF for 24 h before being treated with 100 ng/ml LPS and 1 μM DEX for 4 h. Supernatants were collected, and the levels of glucose and lactate were measured. n = 3; mean ±SEM; * P < 0.05. (D) BMDMs were treated with 25 ng/ml GM-CSF for 24 h before being treated with 100 ng/ml LPS and 1 μM DEX for 4 h. The mRNA levels of GLUT1, HK2 and PFKFB3 were determined by RT-PCR. n = 3; mean ± SEM; * P < 0.05. (E) BMDMs were treated with 25 ng/ml GM-CSF for 24 h before being treated with 100 ng/ml LPS and/or 5 mM ATP and 1 μM DEX for 4 h. Levels of IL-1β, TNF-α and IL-6 were measured by ELISA. n = 3; mean ± SEM; * P < 0.05; ** P < 0.01. (F) BMDMs were treated with 0.5mM or 10mM glucose before being treated with 100 ng/ml LPS and/or 5 mM ATP and 1 μM DEX for 4 h. Levels of IL-1β, TNF-α and IL-6 were measured by ELISA. n = 3; mean ± SEM; * P < 0.05.