Fig. 6

EDH induces autophagy through PERK/eIF2α and IRE1α/XBP-1 s/CHOP pathways in colorectal cancer cells. a The MTT assay was used to evaluate the inhibition rate of colorectal carcinoma cells, which were transfected with PERK or IRE1α siRNAs, followed by incubation with the indicated concentrations of DEH for another 48 h. b The cell activity was detected by colony formation assay. Cells were transfected with PERK or IRE1α siRNAs, followed by incubation with 60 μM DEH for 10 days. The cells were stained with crystal violet staining solution. Scale bar: 200 nm. The number of clones was quantitated and presented to the right of the panel. c The western blotting assay was used to detect the expression of IRE1α, LC3B, and Tubulin. Tubulin was used as an internal control. d Cellular activity was also detected by colony formation assay. The cells were pretreated with 4U8C, followed by incubation with DEH for 10 days. The cells were stained with crystal violet staining solution. Scale bar: 200 nm. The number of clones were quantified and presented below the panel. e The expression of IRE1α, LC3B- II, and Tubulin was detected after DEH treatment with the inhibitor of IRE1α, 4U8C, or DMSO for 48 h. 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