Fig. 3

Enhanced autophagic flux in PDAC cells with Galc variant. a Endogenous activity of GALC in PK59 (Galc variant), HPAFII and Capan-1 (Galc wild-type) was measured using lysosomal GALC analysis. b PK59, HPAF-II and Capan-1 cells were treated with or without 100 μM chloroquine (CQ) for 1 h to monitor autophagy flux. Expression levels of LC3B-I/II in PDAC cell lines was detected in three independent immunoblots. c Autophagy flux unit represents a buildup of LC3B-II with CQ treatment. Densitometric analysis was performed to measure the band intensity of LC3B using ImageJ. d PK59 and HPAF-II cells that stably expressed GFP-LC3 were treated with CQ for 5 h, and GFP-LC3 dots were measured using a fluorescence microscopy. e Number of GFP-LC3 dots were counted manually from the fluorescent images. Indicated numbers above the graphs show an increase in the ratio of LC3 dots under CQ treatment condition relative to that under no treatment condition