Anti sqstm1

Cell or mouse muscle and brain extracts were prepared in lysis buffer containing 50 mM Tris (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1% Triton X-100, halt proteinase inhibitor cocktail (ThermoFisher Scientific) and halt phosphatase inhibitor cocktail (ThermoFisher Scientific), and subjected to western blot analysis with anti-LC3 (Novus Biologicals, NB100-2220), anti-SQSTM1 (Abnova, H00008878-M01), anti-Aβ42 (Invitrogen; 700254), anti-APP (Biolegend; 803001), HRP-conjugated GFP antibody (Santa Cruz Biotechnology, sc9996), anti-HA (Cell Signaling Technology, C29F4), anti-ATG7 (Sigma Aldrich, A2856), anti-LDLR (Abcam, ab52818), anti-LRP1 (Abcam, ab92544), and anti-ACTB/β-actin-HRP (Santa Cruz Biotechnology, sc47778 HRP) antibodies. The band intensity was analyzed using the ImageJ software.

For immunoblot analysis, anti-SQSTM1, anti-LC3 and anti-BECN1 were purchased from BD Biosciences and Sigma Aldrich, respectively. For immunofluorescence studies, anti-LC3 was purchased from Cell Signaling and anti-SQSTM1 from Abnova. Anti-TLR2 was from Genetex and anti-MyD88 was from Cell Signaling. Monoclonal anti- β-actin was provided by Abcam and a polyclonal anti- GAPDH was from Ambion Life Technologies. Antibodies directed against gD, ICP0, phospho-p38α and histone H3 were from Santa Cruz Biotechnology; the antibody against ICP8 was kindly provided by Pr. Bernard Roizman. Horseradish peroxidase anti-rabbit and anti-mouse antibodies were from Santa Cruz Biotechnology. Tetramethyl rhodamine isothiocyanate (TRITC)-conjugated goat anti-rabbit antibody was from Jackson and Alexa Fluor 350 donkey anti-mouse antibody was from Invitrogen. Spautin-1 (10 μM), 3-methyladenin (10 mM), filipin (2.5 μg/ml) and sucrose (0.45 M) were from Sigma Aldrich.