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. 2018 Jan 9;9(8):7902–7917. doi: 10.18632/oncotarget.24083

Figure 3. HPA3P-induced necrosis and morphological changes in human colon cancer cell lines.

Figure 3

All cell lines were treated with HPA3P for 30 min and then stained with Hoechst 33342/PI or PI alone. (A) Pictures of Hoechst 33342/PI-stained cells were taken using an Olympus IX71 fluorescence microscope (magnification LCAch N 20X). (B) PI-stained cells were analysed by flow cytometry. (C) Morphological changes in colon cancer cells treated with HPA3P for 5 min were observed using an Olympus IX71 light microscope (magnification LUCPlaFL N 40X). The black arrows indicate membrane blebs. (D) The cells were incubated with wheat germ agglutinin (WGS) and Hoechst 33342 for 10 min before being washed and then treated HPA3P for 5 min. Morphological changes were observed under an Olympus IX71 fluorescence microscope (magnification LUCPlaFL N 40X). The cell membrane and nucleus were specifically stained by wheat germ agglutinin conjugated with Alexa Fluor 594 (red) and Hoechst 33342 (blue), respectively. The black and white arrows indicate membrane blebs. The LoVo and HT-29 cell lines were treated with 30 μM, and the SW480 and HCT116 p53+/+ cell lines were treated with 50 μM HPA3P.