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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: IκBα targeting promotes oxidative stress-dependent cell death

Fig. 5

Psammaplin-A induces apoptosis via mitochondrial bioenergetics dysfunction. a Growth curves of A549 cells treated with 1 μM cisplatin alone or in combination with 5 μM PSA for 96 h. Data are shown as mean ± sem (n ≥ 3 independent experiments). P-values are from Student’s t-test. **P < 0.01. b Left panel Percentage of apoptotic A549 cells subjected to treatment with 1 μM cisplatin, 5 μM PSA or in combination with two drugs for 48 h, assessed by Annexin V; Right panel: Flow cytometry analysis of Annexine V in A549 cell line. The figure is a representative of three experiments with similar results. Data are shown as mean ± sem (n ≥ 3 independent experiments). P-values are from Student’s t-test. ***P < 0.001,****P < 0.0001. c Flow cytometry experiments measuring intracellular ROS levels in A549 cells treated with 1 μM PSA for 48 h. Data are shown as mean ± sem (n ≥ 3 independent experiments). P-values are from Student’s t-test. **P < 0.01. d Left panel: quantification of mitochondria content; Data are shown as mean ± sem (n ≥ 3 independent experiments). P-values are from Student’s t-test. *P < 0.05. Right panel: The representative images showed that PSA induced extensive mitochondrial fusion. e Representative Western blot showing five ETC proteins (ATP5A, ATP synthase, H+ transporting, mitochondrial F1 complex, α subunit; UQCR2, ubiquinol-cytochrome c reductase core protein II; SDHB, succinate dehydrogenase complex iron sulfur subunit B; COXII, mitochondrially encoded cytochrome c oxidase II; NDUFB8, NADH:ubiquinone oxidoreductase subunit B8) in A549 cells. I, II, III, IV, and V indicate ETC complexes. Vinculin is provided as a loading control. f Oxygen consumption rate (OCR, normalized using CyQUANT fluorescence, arbitrary units, AU). g Mean non-mitochondrial respiration, mean basal respiration (h), maximal respiration (i), proton leak (j) and respiration linked to ATP production (k) in cells treated with vehicle or PSA for 1 day (n ≥ 6). P-values are from Student’s t-test. ****P < 0.0001; ***P < 0.001; **P < 0.01; *P < 0.05

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