Z0034

All rats were perfused using ice-cold 4% PFA, brains were placed in 25% sucrose and cut coronal in a 1:12 series at a thickness of 35 μm. Immunohistochemistry was performed as stated in (Grealish et al., 2015 ). Primary antibodies against hNCAM (Santa Cruz, AB106, monoclonal, mouse, 1:1000); Ox42(CD11b) (Serotec, MCA275G, monoclonal, mouse, 1:1000), GFAP (DAKO, Z0034, polyclonal, rabbit, 1:1000), CD4 (Abcam, AB33775, monoclonal, mouse, 1:1000), CD8 (Serotec, MCA48G, monoclonal, mouse 1:1000), Ox6 (Abcam, AB23990, monoclonal, mouse, 1:500), Ox18 (Abcam, AB33752, monoclonal, mouse, 1:500), HuNu (Chemicon, MAB1281/87, monoclonal, mouse, 1:500), GFP (Abcam, AB13970, polyclonal, chicken, 1:1000), Cre-recombinase (BioLegend, PRB-106P, polyclonal, rabbit, 1:1000) were used.

Double label peroxidase free‐floating immunohistochemistry was undertaken to verify the percentage of neurons and astrocytes targeted with the rAAV‐CBA‐Cre vector. In brief, sections were first incubated in rabbit polyclonal antisera directed against Cre recombinase. Immunoreactivity was revealed with nickel‐enhanced DAB using 1% hydrogen peroxidase which resulted in a black precipitate within the nucleus of the labelled cell. Sections were further incubated in rabbit polyclonal antisera against glial fibrillary acidic protein (GFAP; 1:500; #Z0034, DAKO) or in mouse monoclonal antisera against neuronal nuclei (NeuN; 1:500, #MAB377, Millipore) using the same protocol described above for single label immunohistochemistry and processed using DAB only as a chromogen. The percentage of GFAP‐ and NeuN‐positive cells expressing Cre recombinase was determined with unbiased stereology using the Visiopharm VIS software.