incubated in methanol to facilitate cell permeabilization, in hydrogen peroxide to block endogenous peroxidase activity and in a blocking buffer of Normal Goat Serum (Vector Labs) +0.1%Triton-X in PBS (PBSTx) to inhibit non-specific binding. Next, sections were incubated overnight at RT with an anti-rabbit phosphor-Tau-Ser199,202 polyclonal primary antibody (1:500; Invitrogen, #44–768G). The following day, sections were washed and incubated at RT with an Alexa Fluor® 488 AffiniPure Goat Anti-Rabbit IgG (1:200) secondary antibody prior to slide mounting. Fluorescence was visualized with an Olympus BX51 microscope.
