Fig. 3

The production of IL-1β via IL-10 and STAT-3. a, b STAT-1 and STAT-3 phosphorylation in microglia. Microglial cells from epileptic-seizure mice were incubated for 8 h with or without LPS, and phosphorylated STAT-1 (a) and STAT-3 (b) in cell lysates were measured by western blot (n = 6). The results are representative of three independent experiments. c, d Microglial cells from epileptic-seizure mice were incubated for 8 h with STAT-1 and STAT-3 siRNAs. Then, LPS was added to activate the microglial cells. Western blot analysis showing a dose-dependent reduction of pSTAT-1 (c) and pSTAT-3 (d) signals in cell lysates (n = 6). The graphs show the ratios of pSTAT-1 or pSTAT-3 over GAPDH. Data indicate mean ± SEM of three individual experiments. GAPDH served as an internal control. e The production of IL-1β was assessed by ELISA in supernatants of microglia from epileptic-seizure mice after STAT-1 or STAT-3 knockdown and stimulation with LPS for 12 h (n = 6). Data represent mean ± SEM from three independent experiments.*P < 0.05. f IL-1β-positive cells were evaluated by flow cytometry in microglia from epileptic-seizure mice after LPS stimulation for 8 h in the presence or absence of IL-10 (n = 6). Numbers represent percentages of cells in various tests. g Data represent mean ± SEM from four independent experiments. *P < 0.05; **P < 0.01