Mouse antiluciferase

Brain sections were post-fixed with 4% PFA for 10 minutes and then washed with PBS three times and incubated 30 minutes with blocking solution (PBS/0.1% Triton X-100 containing 10% normal goat serum (Sigma-Aldrich)) and then incubation overnight at 4°C in blocking solution with primary antibodies: mouse anti-CRE (Sigma, F3165–2MG, 1:1000); mouse anti-Luciferase (Sigma, F3165–2MG, 1:1000); mouse anti-Parvalbumin (Sigma, F3165–2MG, 1:1000); mouse anti-MAP2 (Sigma, F3165–2MG, 1:1000); rabbit anti-NeuN (Sigma, F3165–2MG, 1:1000); rabbit anti-IBA1 (Sigma, F3165–2MG, 1:1000). Sections were washed with PBS and incubated for 2h at RT with the secondary antibodies: goat anti-mouse IgG Alexa Fluor 488 (Thermo Fisher, A31560, 1:500) and goat anti-rabbit IgG Alexa Fluor 647 (Invitrogen, A32728, 1:1000) diluted in blocking solution. The sections were washed with PBS and incubated during 10 minutes with DAPI (1:5,000; Sigma), washed, and mounted with Vectashield Antifade Mounting Medium (Vector Labs, H-1000). Immunofluorescence was visualized and imaged with a Keyence BZ-X810 microscope, a Zeiss LSM 510 Meta confocal microscope (Carl Zeiss MicroImaging), equipped with EC Plan-Neofluar 40x/1.30 Oil DIC M27 (420462–9900) and Plan-Apochromat 63x/1.40 Oil DIC M27 (420782–9900) objectives and LSM Image software.