Anti atg5 a0731

The following primary antibodies were used in this study: anti-LRRK2 (MJFF2 [c41-2]; Abcam), anti-human LAMP1 (D2D11; Cell Signaling Technology), anti-mouse LAMP1 (1D4B; Bio-Rad), anti-LC3 (PM036; MBL), anti-LC3 (L7543; Sigma-Aldrich), anti-GFP (A-11122; Invitrogen), anti-p62 (61832; BD Pharmingen), anti-galectin-3 (M3/38; BioLegend), anti-α-tubulin (DM1A; Sigma-Aldrich), anti-CatD (ERP3057Y; ab75852), anti-CatB (D1C7Y; Cell Signaling Technology), anti-Rab10 (D36C4; Cell Signaling Technology), anti-phospho-Thr73 Rab10 (MJF-R21 [ab230261]; Abcam), anti-ATG16L1 (D6D5; CST), anti-ATG5 (A0731; Sigma-Aldrich), anti-FIP200 (17250-1-AP; Proteintech), anti-ATG13 (SAB4200100; Sigma-Aldrich), and anti-Rab29 (MJF-R30-124; Abcam). The following reagents were used at final concentrations as indicated: CQ (50–100 μM; Sigma-Aldrich), LLOMe (1 mM; Sigma-Aldrich), and zymosan (Sigma-Aldrich).

Cells grown in 6‐well format were washed in ice‐cold PBS and scraped in 100 μl of Lysis Buffer (20 mM HEPES pH 7.4, 150 mM NaCl, 1% Triton X‐100, 5 mM EDTA and protease inhibitors) at the relevant time. Protein concentration was estimated using 660 nm Protein Assay Reagent (Pierce) and 20–30 μg of protein loaded on SDS–PAGE gel. Antibodies used were anti‐TECPR1 (8097, Cell Signalling), anti‐ATG5 (A0731, Sigma), anti‐ATG16L1 (8089, Cell Signalling), anti‐GFP (JL8, Clontech) and anti‐β Actin (ab8227, Abcam).