Anti map2

Cell lysates were prepared with Laemmli buffer supplemented with protease inhibitors (complete tablet; Roche) and phosphatase inhibitors (PhosSTOP tablet; Roche) and analysed by Western blotting with the following primary antibodies: anti-γH2AX (Abcam); anti-PTB (Abcam); anti-nPTB (Abcam); anti-βIII Tubulin (Abcam); anti-Tyrosine Hydroxylase (Abcam); anti-GAPDH (Thermo Scientific); anti-MAP2 (Novusbio); anti-DROSHA (Cell Signaling); anti-DICER (Sigma); anti-H2AX (Millipore). Primary antibodies were revealed with peroxidase-conjugated goat anti-mouse or anti-rabbit antibodies (Jackson Immunoresearch Laboratories) and enhanced chemiluminescence system (Super Signal West Pico Pierce or Super Signal West Dura Extended). βIII Tubulin antibody was revealed with alkaline phosphatase-conjugated goat anti-chicken antibody (Santa Cruz) with a colorimetric assay.

Primary antibodies used for immunoblotting and immunofluorescence were as follows: polyclonal anti-prion antibody ab3531 (#ab3531, Abcam, Cambridge, MA), prion protein monoclonal antibody clone SAF-84 (#189775, Cayman, Ann Arbor, MI), recombinant Anti-PrP Fab HuM-D18 (InPro Biotechnology, San Francisco, CA), polyclonal anti-Iba1 (#013-27691, FUJIFILM Wako Chemicals, Richmond, VA), polyclonal anti-GFAP (#AB5541, Millipore Sigma), polyclonal anti-MAP2 (#NB300-213, Novus Biologicals, Centennial, CO). The secondary antibodies used for immunoblotting were goat anti-rabbit IgG-HRP (#474-1506, KPL), goat anti-mouse IgG–HRP (#474-1806, KPL), and goat anti-human IgG, F(ab’)₂ fragment specific, peroxidase-conjugated (#31414, Pierce, Rockford, IL). The secondary antibodies for immunofluorescence were goat anti-mouse IgG antibody (Alexa Fluor 546, #A-11003, ThermoFisher Scientific), donkey anti-rabbit IgG antibody (Alexa Fluor 488, #A-21206, ThermoFisher Scientific), goat anti-rabbit IgG antibody (Alexa Fluor 546, #A-11010, ThermoFisher Scientific), and goat anti-chicken IgG antibody (Alexa Fluor 488, #A-11039, ThermoFisher Scientific).