For IF AT8 (1:500) and MBP (1:2000 polyclonal antibody, Invitrogen) were incubated overnight, followed by the secondary antibodies that included Alexa Fluor® 488 conjugated Goat anti-mouse (1:1000) and Alexa Fluor 647-conjugated Goat anti-chicken (1:200; both from Abcam). Sections were mounted on glass slides with 4′,6-diamidino-2-phenylindole (DAPI) solution (Vectashield, from Vector laboratories). The omission of the primary antibodies resulted in no staining. The sections were photographed at 40× magnification and the images were used to evaluate the fluorescence intensity of MBP immunostaining in regions of interest located in the corpus callosum and stratum alveus. Images were obtained using the laser Olympus FV 1000 D microscope and IF was measure with Fluoview FV1000 software. P-tau was evaluated by calculating the total number cells immuno-stained with a signal larger than 8 microns. All analyses were performed per mouse and for each mouse three slides were measured (Nguyen and Nguyen, 2013 ).
As tMCAo can cause changes in the posterior circulation as well (Lee et al., 2014 (link)), we examined the hippocampal formation and the ipsilateral hemisphere.
As tMCAo can cause changes in the posterior circulation as well (Lee et al., 2014 (link)), we examined the hippocampal formation and the ipsilateral hemisphere.
