Fig. 5

The differential and brain region-dependent effects of long-term peripheral myeloid cell engraftment on astrocytic and neuronal properties. a Representative whole brain images of GFP (green) and common astrocyte marker glial fibrillary acidic protein (GFAP, red) cell deposition in monocyte-engrafted mice. b–c Representative immunofluorescence images of different astrocyte subtypes stained for GFAP (red) and S100β (blue) in the in the cortex (B) and hippocampus (C) of control (WT CON), irradiated (GFP-BM CON), and monocyte-engrafted (GFP-BM REPOP) mice. GFP (green) stains BM-derived cells. d–g Quantification of S100β⁺ and GFAP⁺ cell number in the cortex (D, F) and cell number in the hippocampus (e, g). h Quantification of GFAP staining intensity in the hippocampus. (i) Quantification of NeuN⁺ cell number in the whole cortex. (j) Representative immunofluorescence images stained for neuronal marker NeuN (red) and GFP (green, BM-derived cells) in the cortex and hippocampus. (k) Representative immunofluorescence images of microtubule associated protein 2 (MAP2, red) and GFP (green, BM-derived cells) in the cortex and hippocampus. (l) Representative immunofluorescence brain images of synaptic marker PSD95 (postsynaptic density protein 95, red) and GFP (green, BM-derived cells) in the cortex and hippocampus. L’ provides representative images of spots modeling using Imaris spots module software overlaid on PSD95⁺ staining. (m) Representative immunofluorescence brain images of synaptic marker SV2A (synaptic vesicle glycoprotein 2A, red) and GFP (green, BM-derived cells) in the cortex. (n–s) Quantification of the integrated density of MAP2 staining in the cortex (n) and hippocampus (o), total number of PSD95⁺ spots per field of view in the cortex (p) and hippocampus (q), and total number of SV2A⁺ spots per field of view in the cortex (r) and hippocampus (s). Data are represented as mean ± SEM (n = 4–9). *p < 0.05, **p < 0.01. ***p < 0.001. CTX cortex, HC hippocampus, GFP green fluorescent protein, IBA1 ionized calcium adapter molecule 1. Scale bar ~ 150 μm (B–C, j–left panel); ~ 200 μm (j–right panel); ~40 μm (k); ~ 30 μm (l, m) ~ 15 μm (L’)