Journal: Scientific Reports

Article Title: The novel drug candidate S2/IAPinh improves survival in models of pancreatic and ovarian cancer

doi: 10.1038/s41598-024-56928-z

Figure Lengend Snippet: S2/IAPinh induces activation of the extrinsic apoptosis pathway via degradation of cIAP-1. ( A ) Schematic diagram of the intrinsic and extrinsic apoptosis pathways: arrows represent stimulation. TNFR, tumor necrosis factor receptor; DR4-5, death receptor 4–5; cIAP-1/2, cellular inhibitor of apoptosis protein-1/2; RIPK1, Receptor-interacting serine/threonine-protein kinase 1. ( B ) Protein expression of cIAP-1, cIAP-2, and XIAP in HPAC cells treated with vehicle, SW43 (10 µM), IAPinh (10 µM), or S2/IAPinh (10 µM) for 6 h. The precursor and cleaved forms of caspases 3, 8, and 9 were also analyzed for these cells using Wes automated capillary blotting system (Protein Simple). ( C ) Quantification of protein expression. Relative densitometry of each band normalized to the total protein. Data shown as means ± SEM. ** P < 0.01, **** P < 0.0001. ( D ) Ratio of Caspase 3/7 counts to NucRed counts in HPAC cells treated with vehicle, SW43 (10 µM), IAPinh (10 µM), combination of SW43 (10 µM) and IAPinh (10 µM), or S2/IAPinh (10 µM) measured by the IncuCyte system (Sartorius). Bar graph shows the ratio of Caspase 3/7 at 48 h for each treatment. Data shown as means ± SEM. **** P < 0.001. ( E ) Activity of cell death in AsPC-1 cells was measured using YOYO-1 iodide on the IncuCyte (Sartorius). Representative images of AsPC-1 cells treated with or without Z-VAD-FMK and S2/IAPinh (10 uM) at baseline and 36 h after treatment. Scale bars are equal to 20 µm. ( F ) The AUC of lethal fraction at 36 h. Data shown as means ± SEM. **** P < 0.0001.

Article Snippet: Primary antibodies and dilutions used for the analyses were as follows: cIAP-1 1:2000 for human samples (MAB818, Bio-Techne, Minneapolis, MN), cIAP-1 1:100 for mouse samples (PA5-87514, Invitrogen, Life Technologies, Eugene, OR), cIAP-2 1:500 (NBP-27972, Novus Biologicals, Centennial, CO), X-linked inhibitor of apoptosis protein (XIAP) 1:1000 (610762, BD Biosciences, San Jose, CA), Caspase 3 1:500 (19677–1-AP, Proteintech, Rosemont, IL), Caspase 8 1:50 (NB100-56527, Novus Biologicals, Centennial, CO), and Caspase 9 1:300 (10,380–1-AP, Proteintech, Rosemont, IL).

Techniques: Activation Assay, Expressing, Activity Assay

Journal: Scientific Reports

Article Title: The novel drug candidate S2/IAPinh improves survival in models of pancreatic and ovarian cancer

doi: 10.1038/s41598-024-56928-z

Figure Lengend Snippet: S2/IAPinh induces tumor cell death via activation of the extrinsic apoptosis pathway in vivo. KP2 tumor samples collected from animals at 48 h after the treatment with either vehicle, SW43, IAPinh, or S2/IAPinh, were used for analyses. ( A ) Representative images of TUNEL labeled apoptosis cells in KP2 tumor samples of each group. Nuclei were stained in blue with Hoechst, TUNEL positive cells are in red. Scale bars are equal to 20 µm. ( B ) Quantification of TUNEL positive cells per area in each group. Data are shown as means ± SEM; **** P < 0.0001. ( C ) Protein expression of cIAP-1, cIAP-2, and XIAP in KP2 tumor samples of each group. The precursor and cleaved forms of caspases 3, 8, and 9 were also analyzed for these cells using Wes automated capillary blotting system (Protein Simple). ( D ) Quantification of protein expression. Relative densitometry of each band normalized to the total protein. Data shown as means ± SEM. * P < 0.05, ** P < 0.01, **** P < 0.0001.

Article Snippet: Primary antibodies and dilutions used for the analyses were as follows: cIAP-1 1:2000 for human samples (MAB818, Bio-Techne, Minneapolis, MN), cIAP-1 1:100 for mouse samples (PA5-87514, Invitrogen, Life Technologies, Eugene, OR), cIAP-2 1:500 (NBP-27972, Novus Biologicals, Centennial, CO), X-linked inhibitor of apoptosis protein (XIAP) 1:1000 (610762, BD Biosciences, San Jose, CA), Caspase 3 1:500 (19677–1-AP, Proteintech, Rosemont, IL), Caspase 8 1:50 (NB100-56527, Novus Biologicals, Centennial, CO), and Caspase 9 1:300 (10,380–1-AP, Proteintech, Rosemont, IL).

Techniques: Activation Assay, In Vivo, TUNEL Assay, Labeling, Staining, Expressing