Staining was performed on free-floating brain sections as previously described [37 (link)]. Sections were blocked with 0.2% (v/v) Triton X-100 and 5% (v/v) fetal bovine serum in PBS, and incubated overnight with mouse anti-PV (Millipore, Billerica, MA, MAB 1572; 1:1000) for PV staining or 6E10 (Signet, Dedham, MA; 1:800) for Aβ staining. PV and Aβ staining were visualized using anti-mouse Alex568-labeled secondary antibodies (Invitrogen, Carlsbad, CA; 1:400), incubated for 2 h at RT. Sections were washed and coverslipped in Vectashield including DAPI as a nuclear dye (Vector Laboratories, Burlingame, CA). PV staining was quantified using ImageJ v1.48. PV- and mCherrry-positive cells were counted using image thresholding and automated particle analysis in ImageJ (v1.48). All images were acquired on a Leica DM5000 fluorescence microscope using three different filter cubes (L5, TX2, and Y5) and with a DFC360FX camera (12 bits resolution). For each experiment, identical objective, exposure time, gain settings, and camera settings were used for all images.
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