Fig. 4

Pro-inflammatory mediators are enriched in MP following lipopolysaccharide stimulation of BV2 microglia. a Calcein AM-stained BV2 microglia were stimulated with LPS (20 ng/ml) for 24 h, and MP were isolated by differential centrifugation and stained with anti-annexin V, prior to characterization by flow cytometry. When compared to MP levels in control BV2 microglia, LPS stimulation significantly increased the number of annexin V/calcein AM-positive MP (***p < 0.001 vs control; Student’s t test; n = 4/group). b IL-1β protein expression in enriched MP vs cell lysates of control and LPS-stimulated BV2 microglia. Western blot analysis demonstrated that IL-1β protein was significantly increased in enriched MP following LPS stimulation (^^^p < 0.001 vs control MP; one-way ANOVA with Student-Newman-Keuls correction for multiple comparisons; n = 3/group). Lanes 1, 2, and 3 in both control and LPS refer to sample replicates. c miR-155 expression in enriched MP vs cell lysates of control and LPS-stimulated BV2 microglia. miR-155 was significantly increased in cell lysates of BV2 microglia following LPS stimulation (*p < 0.05 vs control cells), and its expression was elevated further in enriched MP (^^p < 0.01 vs control MP; one-way ANOVA with Student-Newman-Keuls correction for multiple comparisons; n = 4/group). Bars represent mean ± S.E.M. Data represent results of three independent experiments