Fig. 4.

DEH inhibits autophagic flux in colorectal cancer cells. a Fluorescence imaging photographs of HCT 116 and SW620 cells infected with mRFP-GFP-LC3B recombinant adenovirus. The cells were infected with adenovirus for 24 h and then incubated with DMSO, EBSS, 10 μM CQ, and 60 μM DEH for 48 h. Nuclei were stained with DAPI. Scale bar: 10 μm. The average number of autophagosomes (yellow) and autolysosomes (red) per detected cell was counted. All the data were analyzed using the Unpaired Student’s t-test, and p-values less than 0.05 were considered to be statistically significant. *p < 0.05, **p < 0.01, ***p < 0.001. b Western blot analysis of LC3B-II levels in HCT 116 and SW620 cells incubated with or without 60 μM DEH in the absence or presence of 10 mM 3-MA for 48 h. The western blot analysis of LC3B-II levels in HCT 116 and SW620 cells incubated with or without 60 μM DEH in the absence or presence of 10 μM CQ for 48 h. c Western blot analysis of LC3B-II levels in HCT 116 and SW620 cells incubated in normal medium or EBSS with or without 60 μM DEH for 6 h. Western blot analysis of LC3B-II levels in HCT 116 and SW620 cells incubated with or without 60 μM DEH in the absence or presence of 10 nM Baf A1 for 48 h