Fig. 5

Isolation and quantification of CD45-positive brain cells. (a) (i) Cartoon representing brain cells isolation procedure. (ii) Representative flow cytometry dot plots of live cells isolated from brain (except cerebellum and olfactory bulb (OB) and m/Ch). Cells were isolated and stained with control isotype antibody (APC-labeled rat IgG2b) or with APC-labeled rat anti-mouse CD45. Black box marks CD45⁺ cells. Gating strategy was as presented in additional Fig 2s. (iii) The box and whisker plot shows arithmetic mean ± 25-75 quartile (box) and minimum and maximal values (lines) (n = 10) of the percentage of total CD45⁺ cells isolated from 10 months SAMR1 (green) and SAMP8 (red) brains. (b) (i) Cells were stained with PE-Cy7-labeled rat anti-mouse CD45 together with 488 rat anti-mouse CD11b. CD45^m cells are marked with red box and CD45^h cells with a blue box. (ii) Cells were stained with PE-Cy7 rat anti-mouse CD45 plus APC rat IgG2b as isotype control (a); APC rat anti-mouse CD49d (b) or APC rat anti-mouse P2RY12 (c). Shown are representative dot plots obtained for cells isolated from 10 months SAMP8 animals that were similar to cell preparations from young and old SAMR1 and SAMP8 animals. (iii) Box and whisker plot as in panel A iii (n = 10), representing the percentage of total CD45^h cells isolated from 10 months SAMR1 (green) and SAMP8 (red) brains. (ns) Indicates no statistically difference between specified groups applying statistical methods as in material and methods