Fig. 2.

Neuroinflammation is reduced by a single NBE in old mice. a Schematic of serial sections of 25 μm (denoted by the dashed ovals) that were taken from regions ventral to the dentate gyrus, e.g., where an apparent age-associated increase in relative number of CD68⁺ cells was detected (green: thalamus, blue: midbrain, and orange: zona incerta of the thalamus). b Immunofluorescence was performed to assay for CD68-positive (red) activated microglia in the thalamus/hypothalamus/midbrain regions of brains from mice of each cohort. Representative CD68/Hoechst double-positive cells in the specified areas are shown for YY, OO (isochronic controls), and ONBE mice. Isotype-matched IgG negative controls show the absence of non-specific fluorescence. Scale bar 50 μm. c Quantification of the relative frequency of CD68⁺/Hoechst⁺ activated microglia in the thalamus. Neuroinflammation is substantially reduced in ONBE mice when compared to that in OO mice (***p value < 0.00002). The relative numbers of activated microglia are not significantly different between YY mice and ONBE mice (N.S. p value = 0.27). **p value YY versus OO < 0.003. p values were obtained by two-tailed Student’s t test. N of YY = 4, N of OO = 4, N of ONBE = 7