For turnover assays, ATG16L1 rs2241880 knock-in AGS cell lines were subjected to serum starvation (Earle’s Balanced Salt Solution (Sigma) supplemented with sodium bicarbonate 2.2 g/L) or chloroquine (40uM) (Sigma) treatment, for 6 hours.
Western blot was performed as described previously.64 (link) After incubation with the primary antibodies (GRP78 #3177S, LC3B-I/II #2775S, p62 #5114S; Cell Signaling Technology), membranes were incubated with IRDye antibodies (LI-COR Biosciences, Lincoln, NE, USA) or Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (BioRad). Detection was performed using the Odyssey reader or the ImageQuant LAS 500 (GE Life Sciences, Uppsala, Sweden). See supplement for details.
Western blot was performed as described previously.64 (link) After incubation with the primary antibodies (GRP78 #3177S, LC3B-I/II #2775S, p62 #5114S; Cell Signaling Technology), membranes were incubated with IRDye antibodies (LI-COR Biosciences, Lincoln, NE, USA) or Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (BioRad). Detection was performed using the Odyssey reader or the ImageQuant LAS 500 (GE Life Sciences, Uppsala, Sweden). See supplement for details.
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