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. 2018 Aug 6;13(8):e0201920. doi: 10.1371/journal.pone.0201920

Fig 5. Effects of the autophagy and apoptosis inhibitors on the triple treatment-induced growth inhibition in the PANC-1 cells.

Fig 5

The 3-MA and Z-VAD-FMK were used for the autophagy and apoptosis inhibitor analysis, respectively. (A) The PANC-1 cells were pretreated with the autophagy inhibitor 3-MA and the caspase inhibitor Z-VAD-FMK before the triple treatment for 24 h. The result showed that both 3-MA and Z-VAD-FMK could partially enervate the triple treatment-induced growth inhibition in the PANC-1 cells. (B) The pretreatment of Z-VAD-FMK significantly reduced the levels of cleaved caspase-3 and cleaved PARP. The cleaved PARP was quantified relative to the full-length PARP. (C) The pretreatment of 3-MA blocked the conversion of LC3-I to LC3-II. The band intensities of LC3-II and LC3-I were quantified to calculate the LC3-II/LC3-I ratio. β-actin was used as a loading control for each Western blot assay. (D) To better understand the role of autophagy in the triple treatment, 3-MA pretreatment was applied to the combination treatments using US stimulation. (* is used for P < 0.05, ** for P < 0.01, and *** for P < 0.001).