Figure 5. The spleen mediates the enhancing effects of chronic and repeated short-term sleep restriction (CRSR) on LPS-induced blood-brain barrier (BBB) disruption and hippocampal inflammation 24 hours after LPS treatment. Western blot analysis of expression of the tight junction proteins occludin (A) zona occluden-1 (ZO-1; B), and claudin (C) in the brain. (D) Evans blue dye extravasation test. Splenectomy blocked CRSR-mediated exacerbation of LPS-induced decreases in hippocampal tight junction protein levels and increases in Evans blue dye extravasation into the brain. Quantitative real-time PCR (qRT-PCR) analysis of hippocampal TNF-α (E), IL-6 (F), IL-17A (G), and IFN-γ (H) gene expression in each group. Splenectomy blocked CRSR-mediated exacerbation of LPS-induced increases hippocampal expression of these genes. Data represent means ± SEM, n = 6; *P < 0.05, **P < 0.01. N.S., not significant.