568671.fig.002a
(a)
568671.fig.002b
(b)
568671.fig.002c
(c)
568671.fig.002d
(d)
568671.fig.002e
(e)
Figure 2: DHA induces apoptosis and autophagic activation in PC3 and DU145 prostate cancer cells. (a) Cell cycle analysis of PC3 cells exposed to DHA. PC3 cells were incubated with 0−50 μM DHA for 24 h and then subjected to flow cytometry after staining with PI. (b) Increased number of TUNEL-positive cells in DHA-treated cells. PC3 (left) and DU145 (right) cells were incubated with 50 μM DHA for 24 h before staining with TUNEL reagents (scale bar, 200 μm). (c) Levels of cleaved PARP and expression levels of LC3-II in response to DHA. PC3 (left) and DU145 (right) cells were incubated with 0−50 μM DHA for 24 h, and PARP cleavage and the expression levels of LC3-II were monitored by Western blot analysis. (d) Representative images of PC3 (top) and DU145 (bottom) cells transiently expressing GFP-LC3B upon exposure to 50 μM DHA for 2 h (magnification, 600x). (e) Effect of DHA on autophagic flux. PC3 (left) and DU145 (right) cells were incubated with 50 μM DHA and/or 2 μM CQ for 24 h and were then subjected to Western blotting for the detection of LC3-II. Black triangle indicates the lipidated form LC3B (LC3-II), and asterisk indicates nonlipidated form of LC3B (LC3-I).