Fig. 3

25-HC selectively amplifies LPS-induced IL-1β expression and secretion. a-c WT and CH25H KO primary microglia were treated with LPS (0, 0.1, 1, 10, 100 ng/ml) for 24 h. The levels of secreted IL-1β (a), IL-1α (b), and TNFα (c) in the medium were measured by ELISA. d The levels of secreted IL1β from WT and CH25H KO microglia treated with LPS (10 ng/ml) with or without 25-HC (10μg/ml) were measured by ELISA. e Primary microglia were treated with 10 ng/ml LPS in the presence of different concentrations of 25-HC for 24 h. The levels of IL-1β in the media were determined by ELISA and (f) the levels of intracellular pro-IL1β and mature IL1β secreted in the media as measured by Western blotting. Statistical analyses were determined by multiple t test in a, b, c, d; one-way ANOVA in e. *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001. The data shown are representative of three or more independent experiments