Fig. 3

IL-1β and IL1R interact to mediate sclerotic hippocampal antineurogenic effects. A ELISA assay analysis in cultures’ supernatants showed that sclerotic hippocampus derived 3D cultures secrete significantly more IL1-β than cortical counterparts at the indicated time points. B Levels of IL1-β receptor (IL1-R) mRNA are significantly higher in hippocampal cultures. C IL1-R protein is expressed in cell lysates of both hippocampal and cortical 3D cultures. D IL1-R is widely expressed by nestin+ precursor cells in hippocampal 3D cultures. (Scale bar 50 μm). E Blocking endogenous IL1-β by co-treatment with IL-1Ra in 3D hippocampal cultures increases the formation of new neurons (Class III-β-tubulin+EdU+) by 63 ± 16% compared to control cultures. F Significant increase in neurogenesis is also evident by higher levels of the immature (early) neuron proteins NeuroD1 (89% increase) and class III-β-tubulin (74% increase) as measured by western blots. G The proportion of newly formed post-mitotic BrdU+NeuN+ neurons also increased following 3D hippocampal culture treatment with 100 ng/ml of IL-Ra. H In monolayers (2D) cell cultures, addition of exogenous IL1-β causes a significant decrease in density and proportions of new neurons (Class III β-tubulin+EdU+). Addition of the IL-1R blocker IL-1Ra completely blocks this antineurogenic effect. Values are means ± SE. Comparisons made using student’s t test for single comparisons (B, E, F, G) and one (H) or two-way ANOVA (A) with Bonferroni post-hoc for multiple comparisons with *p < 0.05, **p < 0.01, ***p < 0.001 considered significant. Each experiment included at least 12 3D cultures (discs) from 3 different patients