z yvad fmk  (MedChemExpress)

Journal: Journal of Advanced Research

Article Title: Designing an apoptosis reporter by mutagenesis-based insertion of caspase-3 cleavage motif into green fluorescence protein

doi: 10.1016/j.jare.2025.06.070

Figure Lengend Snippet: Monitoring of apoptosis reporter in live cells. (A) HEK293 EGFP #4-8 cells were cultured with or without 1 μM STA, and images were obtained every 30 min for 48 h; scale bar 50 μm. Relative EGFP fluorescence intensity was measured using ImageJ software (N = 3). (B) Protein levels of GFP, caspase-3, cleaved caspase-3, cytochrome C , PARP, and GAPDH by Western blotting in HEK293 EGFP #4-8 cells incubated with STA. Graph, protein levels (N = 3). (C) HEK293 EGFP #4-8 cells were cultured with 1 μM STA co-treated with or without 20 μM Z-DEVD-FMK and 10 μM Z-YVAD-FMK, and images were obtained every 30 min for 48 h. Relative EGFP fluorescence intensity was measured using ImageJ software (N = 3). (D) HEK293 EGFP #4-8 cells were cultured with 100, 200, and 500 μM H 2 O 2 , and images were obtained every 30 min for 24 h. Relative EGFP fluorescence intensity was measured using ImageJ software (N = 3). Error bars are ± SD. *P < 0.05. **P < 0.01. ***P < 0.001. ****P < 0.0001.

Article Snippet: Z-DEVD-FMK (caspase-3 inhibitor, #sc-311558) and Z-YVAD-FMK (caspase-1 inhibitor, #sc-3071) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Cell Culture, Fluorescence, Software, Western Blot, Incubation

Journal: Cell Death Discovery

Article Title: ADA2-deficient cells exhibit increased levels of cell death and metabolic disturbances

doi: 10.1038/s41420-026-03027-9

Figure Lengend Snippet: A Schematic overview of the induction and inhibition of different regulated cell death pathways in vitro. B Cell death in CD14+ monocytes from healthy controls (HC) and DADA2 patients was measured by flow cytometric measurement of staining with Annexin V and Zombie® viability dye. Cells were incubated for 5 h. Apoptosis (TS) and necroptosis (TSZ) were induced by stimulation with 100 nM birinapant (SM) ± 20 µM Z-VAD-FMK (Z-VAD) followed by 20 ng/mL tumour necrosis factor-alpha (TNF-α). Inhibition of necroptosis was achieved by adding 10 µM necrostatin-1s (Nec-1s, TSZN). Plots show median and 25 and 75th percentiles. Data from n = 19 (HC) and n = 14 (DADA2, comprising n = 7 distinct patients) independent experiments are shown. Mann-Whitney-U test, ** p < 0.01. The patients’ phenotype is indicated by colour as displayed in Fig. . Legend: cIAPs cellular inhibitor of apoptosis proteins, DFO deferoxamine, Fer-1 ferrostatin-1, GSDMD gasdermin D, IL-1β interleukin-1beta, MLKL mixed lineage kinase domain like pseudokinase, Nec-1s necrostatin-1s, NSA necrosulfonamide, RIPK1 receptor-interacting serine/threonine-protein kinase 1, RIPK3 receptor-interacting serine/threonine-protein kinase 3, SM Smac mimetic, TRADD tumour necrosis factor receptor type 1-associated DEATH domain protein, TRAF2 TNF receptor-associated factor 2, Z-VAD Z-VAD-FMK, Z-YVAD, Z-YVAD-FMK.

Article Snippet: Prior to induction of cell death, the cells were incubated for 30 min with inhibitors of different forms of RCD as indicated: 20 μM Z-VAD-FMK (#S7023; Selleck Chemicals GmbH, Houston, TX, USA), 20 μM Z-DEVD-FMK (S7312; Selleck Chemicals GmbH), 10 μM necrostatin-1s (#S8641; Selleck Chemicals GmbH), 1 μM necrosulfonamide (#S8251; Selleck Chemicals GmbH), 20 μM Z-YVAD-FMK (#S8507; Selleck Chemicals GmbH), 50 μM deferoxamine (#S5742; Selleck Chemicals GmbH), 2 μM ferrostatin-1 (#S7243; Selleck Chemicals GmbH).

Techniques: Inhibition, In Vitro, Staining, Incubation, MANN-WHITNEY

Journal: Cell Death Discovery

Article Title: ADA2-deficient cells exhibit increased levels of cell death and metabolic disturbances

doi: 10.1038/s41420-026-03027-9

Figure Lengend Snippet: Cell death was measured by flow cytometric measurement of staining with Annexin V and Zombie® viability dye. Apoptosis, necroptosis and pyroptosis were inhibited by incubation with 20 µM Z-VAD-FMK (Z-VAD), 10 µM necrostatin-1s (Nec-1s) and 20 µM Z-YVAD-FMK (Z-YVAD), respectively, for the indicated periods of time. A Data from n = 12 | 12 | 8 (HC) and n = 8 | 8 | 5 (DADA2) independent experiments for Z-VAD|Nec-1s|Z-YVAD, respectively, are shown. B The experiment was performed for n = 2 distinct ADA2 -/- U-937 cell clones. C Data from n = 5 (HC) and n = 4 (DADA2) independent experiments are shown. D Ferroptosis was inhibited by 48-hour incubation with 50 µM deferoxamine (DFO) or 2 µM ferrostatin-1 (Fer-1) in n = 2 HC and DADA2 patients. The median is indicated in all plots. Mann-Whitney-U test, * p < 0.05, ** p < 0.01. The patients’ phenotype is indicated by colour as displayed in Fig. .

Article Snippet: Prior to induction of cell death, the cells were incubated for 30 min with inhibitors of different forms of RCD as indicated: 20 μM Z-VAD-FMK (#S7023; Selleck Chemicals GmbH, Houston, TX, USA), 20 μM Z-DEVD-FMK (S7312; Selleck Chemicals GmbH), 10 μM necrostatin-1s (#S8641; Selleck Chemicals GmbH), 1 μM necrosulfonamide (#S8251; Selleck Chemicals GmbH), 20 μM Z-YVAD-FMK (#S8507; Selleck Chemicals GmbH), 50 μM deferoxamine (#S5742; Selleck Chemicals GmbH), 2 μM ferrostatin-1 (#S7243; Selleck Chemicals GmbH).

Techniques: Staining, Incubation, Clone Assay, MANN-WHITNEY