Fig. 3

Increased cytokine release in WT and Mecp2-null primary mixed glial cell cells challenged with LPS. A multiplex ELISA was conducted on the media collected from the mixed glial cell cultures. At rest (white bars), TNF-α (a), IL-10 (c), and CXCL1 (g) release was increased in Mecp2-null mixed glial culture compared to WT whereas release of IL-1β (b) was decreased. No group differences in IL-6, INF-γ, and IL-12 were observed at rest (d–f). After LPS stimulation (black bars), concentrations of TNF-α (a), IL-10 (c), INF-γ (e), and IL-12 (f) were significantly higher in Mecp2-null compared to WT glial cells. In contrast, blunting of the response to LPS stimulation for IL-1β (b), IL-6 (d), and CXCL1 (g) was observed in Mecp2-null glial cells compared to WT. Across the board, D-NAC 10 μg/mL and D-NAC 100 μg/mL (blue bars) consistently lowered all cytokine levels significantly in WT and Mecp2-null LPS-treated cells (all p ≤ 0.001) with the exception of the chemokine CXCL1 in Mecp2-null glial cells. In most cases, free NAC (red bars) was not effective except for IL-1β in which 10 μg/mL NAC decreased IL-1b levels significantly (*p < 0.05; **p < 0.01; ***p < 0.001)