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. 2016 Oct 18;6:35468. doi: 10.1038/srep35468

Figure 5. APG treatment sensitizes NSCLC cells to TRAIL-induced apoptosis by upregulating DR4 and DR5 levels in a p53-dependent manner.

Figure 5

A549 cells and H1299 cells were treated with APG (20 μM) in the absence or the presence of TRAIL (25 ng/mL) for 24 h. (a) Q-PCR analysis was performed to detect the level of the mRNA transcripts of DR4 and DR5. The results shown are representative of three independent experiments. The histogram shows the mean ± SD. *p < 0.05: significantly different from the control. (b) Western blotting was performed to detect the levels of DR4 and DR5, respectively. (c) Left, A549 cells were treated with APG (20 μM or 40 μM) for the indicated time (4 h or 8 h). The protein levels of p53 in whole cell lysates were determined with specific antibody. Right, A549 cells were treated with APG at different concentrations (0, 10, 20, 40, 80 μM) in the absence or the presence of TRAIL (25 ng/mL) for 24 h. The protein levels of p53 in whole cell lysates were determined with specific antibody. (d) A549 cells were treated with APG or PFT-α at the indicated concentrations for 24 h. DR5 and p53 protein levels in whole cell lysates were determined with specific antibodies. For (b–d), all gels run under the same experimental conditions and the representative images of three different experiments were cropped and shown. Densitometric quantification of the immunoblot data is also shown and data are represented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001. (e) A549 cells were treated with APG (20 μM), TRAIL (25 ng/mL), or the combination of two drugs in the absence or presence of PFT-α (20 μM) for 18 h before determination of cell death by flow cytometry analysis. Data are representative of three independent experiments and are represented as mean ± SD. *p < 0.05.