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caspase 3 inhibitor  (R&D Systems)


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    R&D Systems caspase 3 inhibitor
    HER2-targeted TNFR1-agonists induce TNF-like caspase-1, <t>caspase-3,</t> and caspase-8 activation decoupled from NF-κB signaling in MCF-7 cells (A–C) Caspase-1/3/8 activation of HER2-expressing MCF-7 cells by ICM11 derivatives compared to (rh)TNF. MCF-7 cells were stimulated with increasing compound concentrations for 72 h. Caspase activities were detected intracellularly with FAM-FLICA(R) Caspase 1 Assay Kit (Biomol), CaspaTag Caspase-3 In situ Assay Kit (Merck Millipore), and CaspaTag Caspase-8 In Situ Assay Kit (Merck Millipore). Caspase-1/3/8 activation was normalized to (rh) TNF. (D) NF-κB activation in MCF-7 cells triggered by ICM11 derivatives and (rh) TNF. MCF-7 cells were stimulated with increasing concentrations of ICMs and (rh) TNF for 40 min. NF-κB was stained intracellularly with AF488-labeled anti-NF-κB staining antibody (BD) after lysis, fixation, and permeabilization of cells. (E) Remaining relative cell death of MCF-7 cells after treatment with ICM11 derivatives or (rh)TNF in the presence or absence of caspase inhibitors. MCF-7 cells were incubated with caspase-1 inhibitor (InvivoGen), caspase-3 inhibitor (R&D Systems), caspase-8 inhibitor (InvivoGen), or pan-caspase inhibitor (InvivoGen) at 50 μM and a fixed (rh) TNF or ICM concentration of 5 nM for 72 h. Killing was monitored by green fluorescence signal with SYTOX Green Dead Cell Stain and normalized to (rh) TNF signal. Mean values ±SEM of four independent experiments for each figure are shown. ∗∗∗∗ p < 0.0001,∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05 calculated by utilizing two-way ANOVA multiple analyses and Bonferroni test.
    Caspase 3 Inhibitor, supplied by R&D Systems, used in various techniques. Bioz Stars score: 95/100, based on 412 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/caspase+3+inhibitor+z+devd+fmk/Caspase-3+Inhibitor+Z-DEVD-FMK/pmc13053762-299-20-23
    Average 95 stars, based on 412 article reviews
    caspase 3 inhibitor - by Bioz Stars, 2026-10
    95/100 stars

    Images

    1) Product Images from "Antigen-directed single domain antibody-based TNFR1 agonists elicit preferential killing of HER2-overexpressing cancer cells"

    Article Title: Antigen-directed single domain antibody-based TNFR1 agonists elicit preferential killing of HER2-overexpressing cancer cells

    Journal: iScience

    doi: 10.1016/j.isci.2026.115327

    HER2-targeted TNFR1-agonists induce TNF-like caspase-1, caspase-3, and caspase-8 activation decoupled from NF-κB signaling in MCF-7 cells (A–C) Caspase-1/3/8 activation of HER2-expressing MCF-7 cells by ICM11 derivatives compared to (rh)TNF. MCF-7 cells were stimulated with increasing compound concentrations for 72 h. Caspase activities were detected intracellularly with FAM-FLICA(R) Caspase 1 Assay Kit (Biomol), CaspaTag Caspase-3 In situ Assay Kit (Merck Millipore), and CaspaTag Caspase-8 In Situ Assay Kit (Merck Millipore). Caspase-1/3/8 activation was normalized to (rh) TNF. (D) NF-κB activation in MCF-7 cells triggered by ICM11 derivatives and (rh) TNF. MCF-7 cells were stimulated with increasing concentrations of ICMs and (rh) TNF for 40 min. NF-κB was stained intracellularly with AF488-labeled anti-NF-κB staining antibody (BD) after lysis, fixation, and permeabilization of cells. (E) Remaining relative cell death of MCF-7 cells after treatment with ICM11 derivatives or (rh)TNF in the presence or absence of caspase inhibitors. MCF-7 cells were incubated with caspase-1 inhibitor (InvivoGen), caspase-3 inhibitor (R&D Systems), caspase-8 inhibitor (InvivoGen), or pan-caspase inhibitor (InvivoGen) at 50 μM and a fixed (rh) TNF or ICM concentration of 5 nM for 72 h. Killing was monitored by green fluorescence signal with SYTOX Green Dead Cell Stain and normalized to (rh) TNF signal. Mean values ±SEM of four independent experiments for each figure are shown. ∗∗∗∗ p < 0.0001,∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05 calculated by utilizing two-way ANOVA multiple analyses and Bonferroni test.
    Figure Legend Snippet: HER2-targeted TNFR1-agonists induce TNF-like caspase-1, caspase-3, and caspase-8 activation decoupled from NF-κB signaling in MCF-7 cells (A–C) Caspase-1/3/8 activation of HER2-expressing MCF-7 cells by ICM11 derivatives compared to (rh)TNF. MCF-7 cells were stimulated with increasing compound concentrations for 72 h. Caspase activities were detected intracellularly with FAM-FLICA(R) Caspase 1 Assay Kit (Biomol), CaspaTag Caspase-3 In situ Assay Kit (Merck Millipore), and CaspaTag Caspase-8 In Situ Assay Kit (Merck Millipore). Caspase-1/3/8 activation was normalized to (rh) TNF. (D) NF-κB activation in MCF-7 cells triggered by ICM11 derivatives and (rh) TNF. MCF-7 cells were stimulated with increasing concentrations of ICMs and (rh) TNF for 40 min. NF-κB was stained intracellularly with AF488-labeled anti-NF-κB staining antibody (BD) after lysis, fixation, and permeabilization of cells. (E) Remaining relative cell death of MCF-7 cells after treatment with ICM11 derivatives or (rh)TNF in the presence or absence of caspase inhibitors. MCF-7 cells were incubated with caspase-1 inhibitor (InvivoGen), caspase-3 inhibitor (R&D Systems), caspase-8 inhibitor (InvivoGen), or pan-caspase inhibitor (InvivoGen) at 50 μM and a fixed (rh) TNF or ICM concentration of 5 nM for 72 h. Killing was monitored by green fluorescence signal with SYTOX Green Dead Cell Stain and normalized to (rh) TNF signal. Mean values ±SEM of four independent experiments for each figure are shown. ∗∗∗∗ p < 0.0001,∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05 calculated by utilizing two-way ANOVA multiple analyses and Bonferroni test.

    Techniques Used: Activation Assay, Expressing, In Situ, Staining, Labeling, Lysis, Incubation, Concentration Assay, Fluorescence

    Related Articles

    Saline:

    Article Title: Protection of cells from degeneration and treatment of geographic atrophy
    Article Snippet: In vivo transfection of plasmids coding for two different Alu sequences (pAlu) or empty control vector (pNull) (Bennett et al., 2008; Kaneko et al., 2011; Shaikh et al., 1997) was achieved using 10% Neuroporter (Genlantis). .. Drug treatments. siRNAs formulated in siRNA buffer (20 mMKCL, 0.2 mM MgCl2 in HEPES buffer at pH 7.5; Dharmacon) or phosphate buffered saline (PBS; Sigma-Aldrich); the TLR4 antagonist Ultra Pure Rhodobacter sphaeroides LPS (LPS-RS, InvivoGen), a peptide inhibitor of MyD88 homodimerization IMG-2005 (IMGENEX), control inhibitor (IMGENEX), recombinant IL-18 (Medical & Biological Laboratories), neutralizing rat antibodies against mouse IL-1β (IMGENEX), neutralizing rat antibodies against mouse IL-18 (Medical & Biological Laboratories), isotype control IgGs (R&D Systems or eBioscience as appropriate), Caspase-1 inhibitor Z-WEHD-FMK (R&D Systems), Caspase-3 inhibitor Z-DEVD-FMK (R&D Systems), Caspase control inhibitor Z-FA-FMK (R&D Systems), (2R, 4R)-APDC (Enzo Life Sciences), Glipizide (Sigma-Aldrich), Glyburide (Sigma-Aldrich), and P2X7 receptor antagonist A438079 (Tocris Bioscience) were dissolved in phosphate buffered saline (PBS; Sigma-Aldrich) or dimethyl sulfoxide (DMSO; Sigma-Aldrich), and injected into the vitreous humor in a total volume of 1 μL with a 33-gauge Exmire microsyringe (Ito Corporation). ..

    Control:

    Article Title: Protection of cells from degeneration and treatment of geographic atrophy
    Article Snippet: In vivo transfection of plasmids coding for two different Alu sequences (pAlu) or empty control vector (pNull) (Bennett et al., 2008; Kaneko et al., 2011; Shaikh et al., 1997) was achieved using 10% Neuroporter (Genlantis). .. Drug treatments. siRNAs formulated in siRNA buffer (20 mMKCL, 0.2 mM MgCl2 in HEPES buffer at pH 7.5; Dharmacon) or phosphate buffered saline (PBS; Sigma-Aldrich); the TLR4 antagonist Ultra Pure Rhodobacter sphaeroides LPS (LPS-RS, InvivoGen), a peptide inhibitor of MyD88 homodimerization IMG-2005 (IMGENEX), control inhibitor (IMGENEX), recombinant IL-18 (Medical & Biological Laboratories), neutralizing rat antibodies against mouse IL-1β (IMGENEX), neutralizing rat antibodies against mouse IL-18 (Medical & Biological Laboratories), isotype control IgGs (R&D Systems or eBioscience as appropriate), Caspase-1 inhibitor Z-WEHD-FMK (R&D Systems), Caspase-3 inhibitor Z-DEVD-FMK (R&D Systems), Caspase control inhibitor Z-FA-FMK (R&D Systems), (2R, 4R)-APDC (Enzo Life Sciences), Glipizide (Sigma-Aldrich), Glyburide (Sigma-Aldrich), and P2X7 receptor antagonist A438079 (Tocris Bioscience) were dissolved in phosphate buffered saline (PBS; Sigma-Aldrich) or dimethyl sulfoxide (DMSO; Sigma-Aldrich), and injected into the vitreous humor in a total volume of 1 μL with a 33-gauge Exmire microsyringe (Ito Corporation). ..

    Recombinant:

    Article Title: Protection of cells from degeneration and treatment of geographic atrophy
    Article Snippet: In vivo transfection of plasmids coding for two different Alu sequences (pAlu) or empty control vector (pNull) (Bennett et al., 2008; Kaneko et al., 2011; Shaikh et al., 1997) was achieved using 10% Neuroporter (Genlantis). .. Drug treatments. siRNAs formulated in siRNA buffer (20 mMKCL, 0.2 mM MgCl2 in HEPES buffer at pH 7.5; Dharmacon) or phosphate buffered saline (PBS; Sigma-Aldrich); the TLR4 antagonist Ultra Pure Rhodobacter sphaeroides LPS (LPS-RS, InvivoGen), a peptide inhibitor of MyD88 homodimerization IMG-2005 (IMGENEX), control inhibitor (IMGENEX), recombinant IL-18 (Medical & Biological Laboratories), neutralizing rat antibodies against mouse IL-1β (IMGENEX), neutralizing rat antibodies against mouse IL-18 (Medical & Biological Laboratories), isotype control IgGs (R&D Systems or eBioscience as appropriate), Caspase-1 inhibitor Z-WEHD-FMK (R&D Systems), Caspase-3 inhibitor Z-DEVD-FMK (R&D Systems), Caspase control inhibitor Z-FA-FMK (R&D Systems), (2R, 4R)-APDC (Enzo Life Sciences), Glipizide (Sigma-Aldrich), Glyburide (Sigma-Aldrich), and P2X7 receptor antagonist A438079 (Tocris Bioscience) were dissolved in phosphate buffered saline (PBS; Sigma-Aldrich) or dimethyl sulfoxide (DMSO; Sigma-Aldrich), and injected into the vitreous humor in a total volume of 1 μL with a 33-gauge Exmire microsyringe (Ito Corporation). ..

    Injection:

    Article Title: Protection of cells from degeneration and treatment of geographic atrophy
    Article Snippet: In vivo transfection of plasmids coding for two different Alu sequences (pAlu) or empty control vector (pNull) (Bennett et al., 2008; Kaneko et al., 2011; Shaikh et al., 1997) was achieved using 10% Neuroporter (Genlantis). .. Drug treatments. siRNAs formulated in siRNA buffer (20 mMKCL, 0.2 mM MgCl2 in HEPES buffer at pH 7.5; Dharmacon) or phosphate buffered saline (PBS; Sigma-Aldrich); the TLR4 antagonist Ultra Pure Rhodobacter sphaeroides LPS (LPS-RS, InvivoGen), a peptide inhibitor of MyD88 homodimerization IMG-2005 (IMGENEX), control inhibitor (IMGENEX), recombinant IL-18 (Medical & Biological Laboratories), neutralizing rat antibodies against mouse IL-1β (IMGENEX), neutralizing rat antibodies against mouse IL-18 (Medical & Biological Laboratories), isotype control IgGs (R&D Systems or eBioscience as appropriate), Caspase-1 inhibitor Z-WEHD-FMK (R&D Systems), Caspase-3 inhibitor Z-DEVD-FMK (R&D Systems), Caspase control inhibitor Z-FA-FMK (R&D Systems), (2R, 4R)-APDC (Enzo Life Sciences), Glipizide (Sigma-Aldrich), Glyburide (Sigma-Aldrich), and P2X7 receptor antagonist A438079 (Tocris Bioscience) were dissolved in phosphate buffered saline (PBS; Sigma-Aldrich) or dimethyl sulfoxide (DMSO; Sigma-Aldrich), and injected into the vitreous humor in a total volume of 1 μL with a 33-gauge Exmire microsyringe (Ito Corporation). ..

    Article Title: Dengue Envelope Protein as a Cytotoxic Factor Inducing Hemorrhage and Endothelial Cell Death in Mice
    Article Snippet: The tissue sections were mounted with an aqueous mounting medium (Abcam, Cambridge, MA, USA) and analyzed using a Nikon A1+ confocal microscope (Nikon, Minato, Tokyo, Japan). .. Inhibitors and drugs, such as caspase-3 inhibitor Z-DEVD-FMK (R&D Systems, 10 mg/kg), CSB (a glycosaminoglycan that competes rEIII-cell surface glycoprotein binding; Sigma-Aldrich, 0.5 mg/kg), and NAC (a scavenger of ROS; Sigma-Aldrich, 50 mg/kg), were subcutaneously administered 10 min prior to the rEIII injection injections, following described methods [ ]. ..

    Binding Assay:

    Article Title: Dengue Envelope Protein as a Cytotoxic Factor Inducing Hemorrhage and Endothelial Cell Death in Mice
    Article Snippet: The tissue sections were mounted with an aqueous mounting medium (Abcam, Cambridge, MA, USA) and analyzed using a Nikon A1+ confocal microscope (Nikon, Minato, Tokyo, Japan). .. Inhibitors and drugs, such as caspase-3 inhibitor Z-DEVD-FMK (R&D Systems, 10 mg/kg), CSB (a glycosaminoglycan that competes rEIII-cell surface glycoprotein binding; Sigma-Aldrich, 0.5 mg/kg), and NAC (a scavenger of ROS; Sigma-Aldrich, 50 mg/kg), were subcutaneously administered 10 min prior to the rEIII injection injections, following described methods [ ]. ..

    other:

    Article Title: Caloric restriction leads to druggable LSD1-dependent cancer stem cells expansion.
    Article Snippet: Details of other chemicals used in this study are Caspase-8 Inhibitor Z-IETD-FMK (R&D System, cat#FMK007), Caspase-3 Inhibitor Z-DEVD-FMK (R&D System, cat#FMK004), Pan Caspase Inhibitor Z-VAD-FMK (R&D System, cat#FMK001), m-TOR inhibitor rapamycin (LC Laboratories, R-5000 Rapamycin), IGF-1R/IR inhibitor OSI-906 (cat# S1091, Selleckchem.com), PI3K inhibitor Ly294002 (cat#9901, Cell Signaling Technology), ERK inhibitor Trametinib (GSK-1120212, cat#A-1258, Active Biochem), cycloheximide (Sigma, cat#C7698), actinomycin D((CAS 50-76-0); Santa Cruz Biotechnology, Inc., cat# sc200906) and TRAIL (ALX-201-073-C02, 3v Chimica Srl).

    Article Title: Caloric restriction leads to druggable LSD1-dependent cancer stem cells expansion
    Article Snippet: Details of other chemicals used in this study are Caspase-8 Inhibitor Z-IETD-FMK (R&D System, cat#FMK007), Caspase-3 Inhibitor Z-DEVD-FMK (R&D System, cat#FMK004), Pan Caspase Inhibitor Z-VAD-FMK (R&D System, cat#FMK001), m-TOR inhibitor rapamycin (LC Laboratories, R-5000 Rapamycin), IGF-1R/IR inhibitor OSI-906 (cat# S1091, Selleckchem.com), PI3K inhibitor Ly294002 (cat#9901, Cell Signaling Technology), ERK inhibitor Trametinib (GSK-1120212, cat#A-1258, Active Biochem), cycloheximide (Sigma, cat#C7698), actinomycin D((CAS 50-76-0); Santa Cruz Biotechnology, Inc., cat# sc200906) and TRAIL (ALX-201-073-C02, 3v Chimica Srl).

    Inhibition:

    Article Title: Inhibition of caspase-8 cascade restrains the osteoclastogenic fate of bone marrow cells.
    Article Snippet: Osteoclasts are multinucleated cells of hematopoietic origin, with a pivotal role in bone development and remodeling.. Failure in osteoclast differentiation and activation leads to various bone disorders; thus, attention has focused on a search of molecules involved in osteoclast regulatory pathways.. Caspase-8 appears to be an interesting candidate for further exploration, due to its potential function in bone development and homeostasis.

    Activity Assay:

    Article Title: Inhibition of caspase-8 cascade restrains the osteoclastogenic fate of bone marrow cells.
    Article Snippet: Osteoclasts are multinucleated cells of hematopoietic origin, with a pivotal role in bone development and remodeling.. Failure in osteoclast differentiation and activation leads to various bone disorders; thus, attention has focused on a search of molecules involved in osteoclast regulatory pathways.. Caspase-8 appears to be an interesting candidate for further exploration, due to its potential function in bone development and homeostasis.



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    HER2-targeted TNFR1-agonists induce TNF-like caspase-1, caspase-3, and caspase-8 activation decoupled from NF-κB signaling in MCF-7 cells (A–C) Caspase-1/3/8 activation of HER2-expressing MCF-7 cells by ICM11 derivatives compared to (rh)TNF. MCF-7 cells were stimulated with increasing compound concentrations for 72 h. Caspase activities were detected intracellularly with FAM-FLICA(R) Caspase 1 Assay Kit (Biomol), CaspaTag Caspase-3 In situ Assay Kit (Merck Millipore), and CaspaTag Caspase-8 In Situ Assay Kit (Merck Millipore). Caspase-1/3/8 activation was normalized to (rh) TNF. (D) NF-κB activation in MCF-7 cells triggered by ICM11 derivatives and (rh) TNF. MCF-7 cells were stimulated with increasing concentrations of ICMs and (rh) TNF for 40 min. NF-κB was stained intracellularly with AF488-labeled anti-NF-κB staining antibody (BD) after lysis, fixation, and permeabilization of cells. (E) Remaining relative cell death of MCF-7 cells after treatment with ICM11 derivatives or (rh)TNF in the presence or absence of caspase inhibitors. MCF-7 cells were incubated with caspase-1 inhibitor (InvivoGen), caspase-3 inhibitor (R&D Systems), caspase-8 inhibitor (InvivoGen), or pan-caspase inhibitor (InvivoGen) at 50 μM and a fixed (rh) TNF or ICM concentration of 5 nM for 72 h. Killing was monitored by green fluorescence signal with SYTOX Green Dead Cell Stain and normalized to (rh) TNF signal. Mean values ±SEM of four independent experiments for each figure are shown. ∗∗∗∗ p < 0.0001,∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05 calculated by utilizing two-way ANOVA multiple analyses and Bonferroni test.

    Journal: iScience

    Article Title: Antigen-directed single domain antibody-based TNFR1 agonists elicit preferential killing of HER2-overexpressing cancer cells

    doi: 10.1016/j.isci.2026.115327

    Figure Lengend Snippet: HER2-targeted TNFR1-agonists induce TNF-like caspase-1, caspase-3, and caspase-8 activation decoupled from NF-κB signaling in MCF-7 cells (A–C) Caspase-1/3/8 activation of HER2-expressing MCF-7 cells by ICM11 derivatives compared to (rh)TNF. MCF-7 cells were stimulated with increasing compound concentrations for 72 h. Caspase activities were detected intracellularly with FAM-FLICA(R) Caspase 1 Assay Kit (Biomol), CaspaTag Caspase-3 In situ Assay Kit (Merck Millipore), and CaspaTag Caspase-8 In Situ Assay Kit (Merck Millipore). Caspase-1/3/8 activation was normalized to (rh) TNF. (D) NF-κB activation in MCF-7 cells triggered by ICM11 derivatives and (rh) TNF. MCF-7 cells were stimulated with increasing concentrations of ICMs and (rh) TNF for 40 min. NF-κB was stained intracellularly with AF488-labeled anti-NF-κB staining antibody (BD) after lysis, fixation, and permeabilization of cells. (E) Remaining relative cell death of MCF-7 cells after treatment with ICM11 derivatives or (rh)TNF in the presence or absence of caspase inhibitors. MCF-7 cells were incubated with caspase-1 inhibitor (InvivoGen), caspase-3 inhibitor (R&D Systems), caspase-8 inhibitor (InvivoGen), or pan-caspase inhibitor (InvivoGen) at 50 μM and a fixed (rh) TNF or ICM concentration of 5 nM for 72 h. Killing was monitored by green fluorescence signal with SYTOX Green Dead Cell Stain and normalized to (rh) TNF signal. Mean values ±SEM of four independent experiments for each figure are shown. ∗∗∗∗ p < 0.0001,∗∗∗ p < 0.001, ∗∗ p < 0.01, and ∗ p < 0.05 calculated by utilizing two-way ANOVA multiple analyses and Bonferroni test.

    Article Snippet: We also set out to investigate the consequences of caspase inhibition on cell death induction, harnessing caspase-1 inhibitor (Ac-YVAD-cmk, InvivoGen), caspase-3 inhibitor (Z-DEVD-FMK, R&D Systems), and caspase-8 inhibitor (Z-IETD-FMK, InvivoGen) as well as pan-caspase inhibitor (zVAD-FMK, InvivoGen).

    Techniques: Activation Assay, Expressing, In Situ, Staining, Labeling, Lysis, Incubation, Concentration Assay, Fluorescence

    IBDV-caused apoptosis via mitochondrial pathway. (A) Caspase-3 levels. (B) Caspase-6 levels. (C) Caspase-7 levels. (D) Caspase-8 levels. (E) Caspase-9 levels. (F) Caspase-10 levels. RT-qPCR analysis was conducted on IBDV-infected DF-1 cells at 8, 16, 24, and 32 hpi, with uninfected control groups included for comparison. Results are presented as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Journal: Frontiers in Microbiology

    Article Title: Chicken caspase-3 promotes IBDV replication via the cleavage of IRF7

    doi: 10.3389/fmicb.2026.1741783

    Figure Lengend Snippet: IBDV-caused apoptosis via mitochondrial pathway. (A) Caspase-3 levels. (B) Caspase-6 levels. (C) Caspase-7 levels. (D) Caspase-8 levels. (E) Caspase-9 levels. (F) Caspase-10 levels. RT-qPCR analysis was conducted on IBDV-infected DF-1 cells at 8, 16, 24, and 32 hpi, with uninfected control groups included for comparison. Results are presented as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Article Snippet: Other reagents were the caspase inhibitor Z-VAD (HY-16658B, MedChemExpress, United States), Caspase-3 inhibitor Z-DEVD (HY-12466, MedChemExpress, United States), SYBR Green qPCR Mix (11201ES, YEASEN, China), Cell Counting Kit-8 (C6005, NCM Biotech, China), Annexin V-FITC Apoptosis Detection Kit (556547, BD Biosciences, United States), Lipo8000 transfection reagent (C0533, Beyotime, China), RIPA buffer (P0013B, Beyotime, China), PBS (C0221A, Beyotime, China), Protease Inhibitor Cocktail (CW2200S, CWBIO, China), 5 × SDS loading buffer (CW0027S, CWBIO, China), TRIzon Reagent (CW0580, CWBIO, China), HiFiScript gDNA Removal RT MasterMix (CW2020M, CWBIO, China), protein A/G agarose (80104G, Invivogen, FR).

    Techniques: Quantitative RT-PCR, Infection, Control, Comparison

    The caspase-3 inhibitor Z-DEVD reduces RNA and protein synthesis of IBDV in DF-1 cells. (A) IBDV-infected DF-1 cells were treated with control (no treatment), Z-DEVD (20 μM), or DMSO, and viral loads were measured at 8, 16, 24, and 32 hpi. (B) DF-1 cells were subjected to immunoblotting at 24 hpi after treatment with control, IBDV infection, or simultaneous IBDV infection and Z-DEVD treatment, using the indicated antibodies. β-actin was monitored as a loading control. (C,D) The gray-scale value ratios of target proteins to β-actin are presented as a measure of protein content. Results are expressed as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Journal: Frontiers in Microbiology

    Article Title: Chicken caspase-3 promotes IBDV replication via the cleavage of IRF7

    doi: 10.3389/fmicb.2026.1741783

    Figure Lengend Snippet: The caspase-3 inhibitor Z-DEVD reduces RNA and protein synthesis of IBDV in DF-1 cells. (A) IBDV-infected DF-1 cells were treated with control (no treatment), Z-DEVD (20 μM), or DMSO, and viral loads were measured at 8, 16, 24, and 32 hpi. (B) DF-1 cells were subjected to immunoblotting at 24 hpi after treatment with control, IBDV infection, or simultaneous IBDV infection and Z-DEVD treatment, using the indicated antibodies. β-actin was monitored as a loading control. (C,D) The gray-scale value ratios of target proteins to β-actin are presented as a measure of protein content. Results are expressed as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Article Snippet: Other reagents were the caspase inhibitor Z-VAD (HY-16658B, MedChemExpress, United States), Caspase-3 inhibitor Z-DEVD (HY-12466, MedChemExpress, United States), SYBR Green qPCR Mix (11201ES, YEASEN, China), Cell Counting Kit-8 (C6005, NCM Biotech, China), Annexin V-FITC Apoptosis Detection Kit (556547, BD Biosciences, United States), Lipo8000 transfection reagent (C0533, Beyotime, China), RIPA buffer (P0013B, Beyotime, China), PBS (C0221A, Beyotime, China), Protease Inhibitor Cocktail (CW2200S, CWBIO, China), 5 × SDS loading buffer (CW0027S, CWBIO, China), TRIzon Reagent (CW0580, CWBIO, China), HiFiScript gDNA Removal RT MasterMix (CW2020M, CWBIO, China), protein A/G agarose (80104G, Invivogen, FR).

    Techniques: Infection, Control, Western Blot

    Overexpression of caspase-3 promotes IBDV replication. DF-1 cells were transfected with either pcDNA3.1 or pcDNA3.1-Caspase-3, and then infected with IBDV at 12 h post-transfection. (A) Viral loads were measured at 8, 16, 24, and 32 hpi. (B) Cells were collected at 24 hpi and analyzed by Western blotting. (C) The gray-scale value ratios of target proteins to β-actin are presented as a measure of protein content. Results are expressed as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Journal: Frontiers in Microbiology

    Article Title: Chicken caspase-3 promotes IBDV replication via the cleavage of IRF7

    doi: 10.3389/fmicb.2026.1741783

    Figure Lengend Snippet: Overexpression of caspase-3 promotes IBDV replication. DF-1 cells were transfected with either pcDNA3.1 or pcDNA3.1-Caspase-3, and then infected with IBDV at 12 h post-transfection. (A) Viral loads were measured at 8, 16, 24, and 32 hpi. (B) Cells were collected at 24 hpi and analyzed by Western blotting. (C) The gray-scale value ratios of target proteins to β-actin are presented as a measure of protein content. Results are expressed as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Article Snippet: Other reagents were the caspase inhibitor Z-VAD (HY-16658B, MedChemExpress, United States), Caspase-3 inhibitor Z-DEVD (HY-12466, MedChemExpress, United States), SYBR Green qPCR Mix (11201ES, YEASEN, China), Cell Counting Kit-8 (C6005, NCM Biotech, China), Annexin V-FITC Apoptosis Detection Kit (556547, BD Biosciences, United States), Lipo8000 transfection reagent (C0533, Beyotime, China), RIPA buffer (P0013B, Beyotime, China), PBS (C0221A, Beyotime, China), Protease Inhibitor Cocktail (CW2200S, CWBIO, China), 5 × SDS loading buffer (CW0027S, CWBIO, China), TRIzon Reagent (CW0580, CWBIO, China), HiFiScript gDNA Removal RT MasterMix (CW2020M, CWBIO, China), protein A/G agarose (80104G, Invivogen, FR).

    Techniques: Over Expression, Transfection, Infection, Western Blot

    Overexpression of caspase-3 inhibits the activation of the IRF7 antiviral pathway and promotes apoptosis induced by IBDV. DF-1 cells were transfected with either pcDNA3.1 or pcDNA3.1-Caspase-3, and then infected with IBDV at 12 h post-transfection. (A,B) Annexin V-FITC and PI flow cytometry were performed to detect the number of apoptotic cells in DF-1 cells at 24 hpi. (C,D) RT-qPCR analysis was conducted at 8, 16, 24, and 32 hpi, with uninfected control groups included for comparison. Results are presented as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Journal: Frontiers in Microbiology

    Article Title: Chicken caspase-3 promotes IBDV replication via the cleavage of IRF7

    doi: 10.3389/fmicb.2026.1741783

    Figure Lengend Snippet: Overexpression of caspase-3 inhibits the activation of the IRF7 antiviral pathway and promotes apoptosis induced by IBDV. DF-1 cells were transfected with either pcDNA3.1 or pcDNA3.1-Caspase-3, and then infected with IBDV at 12 h post-transfection. (A,B) Annexin V-FITC and PI flow cytometry were performed to detect the number of apoptotic cells in DF-1 cells at 24 hpi. (C,D) RT-qPCR analysis was conducted at 8, 16, 24, and 32 hpi, with uninfected control groups included for comparison. Results are presented as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Article Snippet: Other reagents were the caspase inhibitor Z-VAD (HY-16658B, MedChemExpress, United States), Caspase-3 inhibitor Z-DEVD (HY-12466, MedChemExpress, United States), SYBR Green qPCR Mix (11201ES, YEASEN, China), Cell Counting Kit-8 (C6005, NCM Biotech, China), Annexin V-FITC Apoptosis Detection Kit (556547, BD Biosciences, United States), Lipo8000 transfection reagent (C0533, Beyotime, China), RIPA buffer (P0013B, Beyotime, China), PBS (C0221A, Beyotime, China), Protease Inhibitor Cocktail (CW2200S, CWBIO, China), 5 × SDS loading buffer (CW0027S, CWBIO, China), TRIzon Reagent (CW0580, CWBIO, China), HiFiScript gDNA Removal RT MasterMix (CW2020M, CWBIO, China), protein A/G agarose (80104G, Invivogen, FR).

    Techniques: Over Expression, Activation Assay, Transfection, Infection, Flow Cytometry, Quantitative RT-PCR, Control, Comparison

    Caspase-3 cleaves IRF7. (A,B) DF-1 cells were transfected with the HA-tagged IRF7 plasmid and infected with IBDV at 12 h post-transfection. Cells were collected at 12 and 24 hpi for Western blot analysis. (C,D) The indicated plasmids were transformed, purified from BL21, and subjected to in vitro cleavage assays, followed by SDS-PAGE analysis and immunoblotting with the specified antibodies. (E,F) DF-1 cells were co-transfected with Flag-tagged Caspase-3 and HA-tagged IRF7 plasmids, and analyzed by Western blotting at 24 and 36 h post-transfection. (G,H) IBDV-infected DF-1 cells were treated with Z-DEVD (20 μM) for 24 h and the protein level of IRF7 was analyzed by Western blotting. Results are expressed as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Journal: Frontiers in Microbiology

    Article Title: Chicken caspase-3 promotes IBDV replication via the cleavage of IRF7

    doi: 10.3389/fmicb.2026.1741783

    Figure Lengend Snippet: Caspase-3 cleaves IRF7. (A,B) DF-1 cells were transfected with the HA-tagged IRF7 plasmid and infected with IBDV at 12 h post-transfection. Cells were collected at 12 and 24 hpi for Western blot analysis. (C,D) The indicated plasmids were transformed, purified from BL21, and subjected to in vitro cleavage assays, followed by SDS-PAGE analysis and immunoblotting with the specified antibodies. (E,F) DF-1 cells were co-transfected with Flag-tagged Caspase-3 and HA-tagged IRF7 plasmids, and analyzed by Western blotting at 24 and 36 h post-transfection. (G,H) IBDV-infected DF-1 cells were treated with Z-DEVD (20 μM) for 24 h and the protein level of IRF7 was analyzed by Western blotting. Results are expressed as mean ± SD, analyzed using a two-sample Student’s t -test. * P < 0.05, ** P < 0.01.

    Article Snippet: Other reagents were the caspase inhibitor Z-VAD (HY-16658B, MedChemExpress, United States), Caspase-3 inhibitor Z-DEVD (HY-12466, MedChemExpress, United States), SYBR Green qPCR Mix (11201ES, YEASEN, China), Cell Counting Kit-8 (C6005, NCM Biotech, China), Annexin V-FITC Apoptosis Detection Kit (556547, BD Biosciences, United States), Lipo8000 transfection reagent (C0533, Beyotime, China), RIPA buffer (P0013B, Beyotime, China), PBS (C0221A, Beyotime, China), Protease Inhibitor Cocktail (CW2200S, CWBIO, China), 5 × SDS loading buffer (CW0027S, CWBIO, China), TRIzon Reagent (CW0580, CWBIO, China), HiFiScript gDNA Removal RT MasterMix (CW2020M, CWBIO, China), protein A/G agarose (80104G, Invivogen, FR).

    Techniques: Transfection, Plasmid Preparation, Infection, Western Blot, Transformation Assay, Purification, In Vitro, SDS Page

    IBDV VP3 interacts with caspase-3. After co-transfecting DF-1 cells with corresponding plasmids, we harvested samples 48 h later. Cells were lysed and immunoprecipitation was performed using Flag-tag antibodies, followed by Western blotting detection. (A) Flag- Caspase-3 and HA-IRF7. (B) Flag- Caspase-3 and HA-VP3. (C) DF-1 cells transfected with p3 × Flag-CMV-14-Caspase-3 and pcDNA3.1-HA-VP3 were subjected to immunofluorescence staining with anti-Flag rabbit mAb and anti-HA mouse mAb.

    Journal: Frontiers in Microbiology

    Article Title: Chicken caspase-3 promotes IBDV replication via the cleavage of IRF7

    doi: 10.3389/fmicb.2026.1741783

    Figure Lengend Snippet: IBDV VP3 interacts with caspase-3. After co-transfecting DF-1 cells with corresponding plasmids, we harvested samples 48 h later. Cells were lysed and immunoprecipitation was performed using Flag-tag antibodies, followed by Western blotting detection. (A) Flag- Caspase-3 and HA-IRF7. (B) Flag- Caspase-3 and HA-VP3. (C) DF-1 cells transfected with p3 × Flag-CMV-14-Caspase-3 and pcDNA3.1-HA-VP3 were subjected to immunofluorescence staining with anti-Flag rabbit mAb and anti-HA mouse mAb.

    Article Snippet: Other reagents were the caspase inhibitor Z-VAD (HY-16658B, MedChemExpress, United States), Caspase-3 inhibitor Z-DEVD (HY-12466, MedChemExpress, United States), SYBR Green qPCR Mix (11201ES, YEASEN, China), Cell Counting Kit-8 (C6005, NCM Biotech, China), Annexin V-FITC Apoptosis Detection Kit (556547, BD Biosciences, United States), Lipo8000 transfection reagent (C0533, Beyotime, China), RIPA buffer (P0013B, Beyotime, China), PBS (C0221A, Beyotime, China), Protease Inhibitor Cocktail (CW2200S, CWBIO, China), 5 × SDS loading buffer (CW0027S, CWBIO, China), TRIzon Reagent (CW0580, CWBIO, China), HiFiScript gDNA Removal RT MasterMix (CW2020M, CWBIO, China), protein A/G agarose (80104G, Invivogen, FR).

    Techniques: Immunoprecipitation, FLAG-tag, Western Blot, Transfection, Immunofluorescence, Staining