Data represent detection of CCR5 transcripts in sections of mouse cortex through FISH (A–E) and quantitative gene expression following FACS isolation (F and G).

(A) Representative images from healthy mouse cortex show absence of co-localization of CCR5 transcripts (red, column 2) with TUBB3+ve (yellow) neurons (column 3). Insets represent digitally magnified field of view from the image. Inset in column 2 shows DAPI+ve (blue) nuclei and negligible CCR5 (red). Schematic on left represents location of field of view. Scale bar, 50 μm.

(B) Microglia (CX3CR1+ve, yellow) co-localize with CCR5 (red) as seen in column 3 in healthy cortex. Scale bar, 50 μm.

(C) CCR5 expression co-localizes with TUBB3+ve neurons and non-neuronal DAPI+ve nuclei at 12 h after stroke. Scale bar, 100 μm.

(D) Column 1 represents a projection image (3D visualization, Imaris) of neurons at higher magnification (TUBB3+ve, yellow; DAPI+ve, blue) that express CCR5 (red) at 12 h after stroke. Columns 2 and 3 represent images from column 1 that were processed with spot detection feature from Imaris (Bitplane software) as a visual aid for co-localization of TUBB3, CCR5 (column 2) and DAPI (column 3). Scale bar, 10 μm.

(E) CCR5 expression at 7 days after stroke; additional images are in Figure S1 . CCR5 is expressed in neurons at lower abundance in TUBB3+ neurons compared to 12 h and non-neuronal DAPI+ve cells. Columns 2 and 3 were processed similar to (D). Scale bar, 20 μm.

(F) Representative dot plots from FACS show gating strategy used to isolate allophycocyanin (APC)-expressing NCAM+ve neurons isolated from peri-infarct cortex. Similar gating strategy was used for isolation of CD11b+ve cells.

(G) Data on CCR5 gene expression quantified from FACS-isolated cells from naive and 12 h, 7 days,14 days, and 28 days post stroke. Gene expression data are from four different observations for NCAM pre-stroke, NCAM 12 h, CD11b pre-stroke, CD11b 12 h, and three different observations from all other groups. Data are mean ± SEM.