Figure 9. Effect of LC3 knockdown on 15d-PGJ2 induced cytoplasmic vacuolation and cell death.
Clones of HCT116 cells expressing GFPshRNA (CONshRNA) or LC3 shRNA were obtained. A. Western blots of total cell lysates showing the effect of knockdown of LC3 on 15d-PGJ2 induced accumulation of ubiquitinated proteins, expression of ER stress markers, Bip and CHOP and expression and processing of LC3. B. Phase contrast images showing effect of LC3 knockdown on 15d-PGJ2 induced cytoplasmic vacuolation. C. Bar graph showing percentage of vacuolated HCT116 cells that were expressing CONshRNA or LC3 shRNA with or without 15d-PGJ2 treatment. D. Cell viability determined by trypan blue exclusion assay after growing clones of HCT116 cells for 24h in growth medium after incubating with 15d-PGJ2 (20 µM) for 9h. Data represents an average of three independent experiments. E. Western blots of total cell lysates showing effect of LC3 knockdown on 15d-PGJ2 induced expression of p53, p21, p27 in HCT116 cells.