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Arenobufagin induces caspase-dependent apoptosis in non-small-cell lung cancer (NSCLC) cells. ( A ) A549 and NCI-H460 cells were treated with the indicated concentrations of arenobufagin for 24 h, and <t>apoptotic</t> morphological changes were evaluated by Hoechst 33258 staining; ( B , C ) A549 and NCI-H460 cells were treated with arenobufagin at the indicated concentrations for 24 h, and protein extracts were subjected to Western blot assay with indicated antibodies; ( D , E ) A549 cells were pre-treated with the caspase inhibitor Z-VAD (40 μΜ) for 1 h, followed by incubation with arenobufagin (Are, 25 nM) for 24 h; ( D ) poly (ADP-ribose) polymerase (PARP) cleavage was analyzed by Western blotting; ( E ) The cell viability was detected via trypan blue exclusion assay. ** p < 0.01, arenobufagin-treated group versus arenobufagin and Z-VAD combination group.
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Arenobufagin induces caspase-dependent apoptosis in non-small-cell lung cancer (NSCLC) cells. ( A ) A549 and NCI-H460 cells were treated with the indicated concentrations of arenobufagin for 24 h, and apoptotic morphological changes were evaluated by Hoechst 33258 staining; ( B , C ) A549 and NCI-H460 cells were treated with arenobufagin at the indicated concentrations for 24 h, and protein extracts were subjected to Western blot assay with indicated antibodies; ( D , E ) A549 cells were pre-treated with the caspase inhibitor Z-VAD (40 μΜ) for 1 h, followed by incubation with arenobufagin (Are, 25 nM) for 24 h; ( D ) poly (ADP-ribose) polymerase (PARP) cleavage was analyzed by Western blotting; ( E ) The cell viability was detected via trypan blue exclusion assay. ** p < 0.01, arenobufagin-treated group versus arenobufagin and Z-VAD combination group.

Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

Article Title: Arenobufagin Induces Apoptotic Cell Death in Human Non-Small-Cell Lung Cancer Cells via the Noxa-Related Pathway

doi: 10.3390/molecules22091525

Figure Lengend Snippet: Arenobufagin induces caspase-dependent apoptosis in non-small-cell lung cancer (NSCLC) cells. ( A ) A549 and NCI-H460 cells were treated with the indicated concentrations of arenobufagin for 24 h, and apoptotic morphological changes were evaluated by Hoechst 33258 staining; ( B , C ) A549 and NCI-H460 cells were treated with arenobufagin at the indicated concentrations for 24 h, and protein extracts were subjected to Western blot assay with indicated antibodies; ( D , E ) A549 cells were pre-treated with the caspase inhibitor Z-VAD (40 μΜ) for 1 h, followed by incubation with arenobufagin (Are, 25 nM) for 24 h; ( D ) poly (ADP-ribose) polymerase (PARP) cleavage was analyzed by Western blotting; ( E ) The cell viability was detected via trypan blue exclusion assay. ** p < 0.01, arenobufagin-treated group versus arenobufagin and Z-VAD combination group.

Article Snippet: Characteristic apoptotic morphological changes were assessed by Hoechst 33258 staining, using the Hoechst staining kit (Beyotime, Haimen, China).

Techniques: Staining, Western Blot, Incubation, Trypan Blue Exclusion Assay