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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: FGF2/FGFR1 regulates autophagy in FGFR1-amplified non-small cell lung cancer cells

Fig. 6

Inhibition of autophagy by beclin-1 silencing enhances apoptosis after AZD4547 treatment. a H1581 cells were transfected with beclin-1 siRNA or control siRNA using Lipofectamine RNAiMAX transfection reagent. At 24 h after transfection, the cells were treated with or without AZD4547 (1 μM) for 24 h. Cell viability was determined by CCK-8 assay; mean ± SD, n = 3, ***p < 0.001. b H520 cells were treated as described in (a), and cell viability was measured by CCK-8 assay; mean ± SD, n = 3, ***p < 0.001. c and d H1581 and H520 cells transfected with beclin-1 siRNA or control siRNA were treated with or without AZD4547 (1 μM) for another 24 h. Cleavage of both PARP and caspase-9 were analyzed by western blot assay. e and f H1581 and H520 cells transfected with beclin-1 siRNA or control siRNA were treated with or without AZD4547 (1 μM) for another 24 h. The last 5 h was in the presence or absence of z-VAD-fmk (20 μM). PARP cleavage and tubulin were analyzed by western blot assay. g H1581 and H520 cells transfected with beclin-1 siRNA or control siRNA were treated with or without AZD4547 (1 μM) for another 24 h. Cell apoptosis was determined by flow cytometry using FITC Annexin V/PI staining. The horizontal and vertical axes represent labeling with FITC Annexin V/PI, respectively. LR (Q3) represents early apoptotic cells (positive for Annexin V only), UR (Q2) represents late apoptotic cells (positive for both Annexin V and PI), and LL (Q4) represents live cells. h H1581 cells were treated as described in (g upper panel). The number of early/late apoptotic cells is shown as the sum of Annexin V positive and Annexin V/PI double positive cells; mean ± SD, n = 3, ***p < 0.001. i H520 cells were treated as described in (g lower panel); mean ± SD, n = 3, **p < 0.01

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