Research Paper Volume 12, Issue 18 pp 17990—18007

Figure 4. Inhibitory effects of minocycline on inflammatory responses in microglial cells. (A) Adult mice (IMG) and neonatal murine (BV-2) microglia were pretreated with minocycline (20 μM) for 30 min before stimulation with LPS (100 ng·mL⁻¹) for 24 h. The cell culture medium was then harvested to determine the nitrite content by the Griess reaction. IMG (B) and BV-2 (C) microglia were pretreated with minocycline (20 μM) for 30 min before stimulation with LPS (100 ng·mL⁻¹) for 24 h. Whole-cell lysates were subjected to western blot analysis for iNOS and COX-2 expression. (D) Cells were pretreated with minocycline (20 μM) for 30 min before stimulation with LPS (100 ng·mL⁻¹) for 90 min. Whole-cell lysate proteins were subjected to western blot analysis using antibodies against phospho-mTOR or phospho-CREB. Similar results were obtained from at least three independent experiments. * p < 0.05, compared with the control group; # p < 0.05, compared with the only LPS group.