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. 2015 Mar 24;11(2):314–331. doi: 10.1080/15548627.2015.1017182

Figure 8.

Figure 8.

3-AWA-mediated stimulation of autophagy in 3-AWA-resistant, as well as TP53+/+ and TP53−/− cells. (A and B) Microphotograph shows AVO due to acridine orange staining of 3-AWA resistant MiaPaCa-2 cells treated with the indicated concentrations of 3-AWA, rapamycin (100 nM) and 3-AWA plus 3-MA (5 mM) for 12 h to characterize autophagy. Quantification of AO-positive cells from the above experiment was analyzed by fluorescence microscopy. Three random fields representing 100 cells were counted. (C and D) Representative fluorescence microscopy images show that HCT116 TP53+/+ and HCT116 TP53−/− cells were treated with the indicated concentrations of 3-AWA, 100 nM rapamycin, 0.75 μM 3-AWA plus 5 mM 3-MA and vehicle DMSO for 12 h to detect autophagy by acridine orange dye. Quantification of AO-positive cells from the above experiment was analyzed by fluorescence microscopy. Three random fields representing 100 cells were counted. (E) Whole cell lysates prepared from the above treated HCT116 TP53+/+ and HCT116 TP53−/− cells were immunoblotted and probed with LC3B-I/II, SQSTM-1 and loading control ACTB. The data represent the mean value ± SE of 3 independent experiments, **P < 0.01.