Activated Eif2b5^R132H/R132H primary microglial cells exhibit impaired production of cytokines.

<p>Primary microglial cells were isolated from Eif2b5^R132H/R132H (Mut) and wild-type (WT) mice. (A) Representative analysis of PE-labelled anti-CD11b staining followed by flow cytometry analysis. Both Mut and WT cultures were 92–95% CD11b-positive (white area). (B,C,D) Both cultures were treated with 0.1 µg/ml LPS for the indicated times followed by media collection and cell harvest for RNA and protein extraction. qRT-PCR of IL-6, TNF-α and IL-1β mRNA levels is shown in (B), bars represent the average of 3 independent assays, normalised to WT cells at 3 h of LPS treatment ± SEM. Intracellular IL-6 and IL-1β protein levels were determined by immunoblot analyses with β-actin as loading control, representative blots of 3 independent experiments are shown in (C); protein concentrations of IL-6 and TNF-α in the media was measured by ELISA and a representative of 3 independent experiments is shown in (D).