Fig. 3
ASPP2 negatively regulate basal autophagy in pancreatic cancer cell lines. a Immunoblotting analysis was performed to determine LC3-II in different pancreatic cancer cell lines; b GFP-LC3 was transfected into PC cells with different expression levels of ASPP2 and autophagosome formations was observed under microscopy. Representative picture and the percentage of autophagic cells were calculated in 5 random fields. c Western blot assay was performed to determine the expression of autophagy-related protein in BxPC-3 cells with stable ASPP2 and SW1990 cells with ASPP2 stably knockdown. d The cells in (C) were transfected with GFP-LC3 expression plasmids and autophagosome formations was observed under microscopy. e Representative electron micrographs of autophagic vesicles were shown. Arrows denote autophagosomes. Magnified images also were shown. f Representative images of LC3 staining in sh-con/sh-ASPP2 SW1990 cells infected with adenovirus-delivering mRFP-GFP-LC3. Quantification of autophagosome and autolysosome formation represents punctures staining sites per cell of 5 independent images. Data represent the mean ± SD of three independent experiments (*P < 0.05, **P < 0.01)