Anti ndp52

Anti-S. pneumoniae (SSI), anti-GFP (Cell Signaling), anti-Myc (9B11, Cell Signaling), anti-Flag (Wako), anti-Galectin-3 (SINO BIOLOGICAL), anti-Calcoco2 (Proteintech for WB), anti-NDP52 (Gene Tex (GTX115378) for IF), anti-K63 linked Ub (clone Apu3, EMD Millipore), anti-LC3, p62, RFP, Atg16L1, ubiquitin (MBL), and anti-actin (Santa Cruz Biotechnology, Inc.) were used as primary antibodies. An HRP-conjugated goat anti-rabbit or anti-mouse antibodies (Jackson Laboratories) were used as secondary antibodies for immunoblotting. FITC- or TRITC-conjugated goat anti-rabbit or anti-mouse IgG antibodies (Sigma-Aldrich) were used as secondary antibodies for immunostaining. DAPI (4′,6-diamidino- 2-phenylindole, Sigma-Aldrich) was used for DNA staining. LysoTracker DND-99 was purchased from molecular probes. 10 µM rapamycin and 40 µM chloroquine (Selleck chemical), and 30 µM PYR-41 (UBPBio), and 10 mM 3-methyladenine (3-MA, Wako) were used as autophagy inducer or inhibitor. 300 µM Apocynin (Selleck), 10 mM GSH (Cayman Chemical), 2.5 mM NAC (Sigma-Aldrich) were used as antioxidative reagents. All other reagents were purchased from Sigma-Aldrich. All antibodies were used at 1:100 for immunofluorescence staining and 1:1000 for western blotting.

The drugs used in this study were as follows: picrotoxin (Sigma), a GABA-receptor inhibitor; two different autophagy inhibitors, 3MA (3-methyladenine; Santa Cruz Biotechnology) and Bafilomycin (Bafilomycin A1; AdipoGen); and the proteasome inhibitor MG132 (Chemscene). The antibodies used were as follows: anti-GluR2 (Millipore), anti-LC3 (M152–3; MLB), A0024 (Daco), T22 (Millipore), Tau5 (Millipore), anti-pS396-tau (Invitrogen), anti-NDP52 (GeneTex), Alexa Fluor 488– and Alexa Fluor 568–conjugated secondary antibodies (Invitrogen), gold-conjugated (5 nm) secondary antibodies (British BioCell International), and horseradish peroxidase–conjugated secondary antibodies (Jackson ImmunoResearch Laboratories).