Figure 6. The spleen mediated the enhancing effects of chronic and repeated short-term sleep restriction (CRSR) on lipopolysaccharide (LPS)-induced transition of microglia to M1 phenotype 24 hours after LPS treatment. (A) Western blot analysis of hippocampal ionized calcium-binding adapter molecule 1 (Iba1) expression in each group. Splenectomy blocked CRSR-mediated exacerbation of the LPS-induced increase in hippocampal Iba1 expression. Quantitative real-time PCR (qRT-PCR) analysis of hippocampal expression of the microglial M1 markers CD16 (B), CD11b (C), CD32 (D), and iNOS (E) and the M2 markers Arg-1 (F), TGF-β (G), CD206 (H), and YM1 (I). Splenectomy blocked CRSR-mediated exacerbation of LPS-induced increases in microglial M1 markers and decreases in M2 markers in the hippocampus. Data represent means ± SEM, n = 6; *P < 0.05, **P < 0.01. N.S., not significant.