anti active caspase 3  (R&D Systems)

Journal: Nature Genetics

Article Title: Single-cell spatial transcriptomic analysis of human skin anatomy

doi: 10.1038/s41588-026-02552-8

Figure Lengend Snippet: a , Schematic of human skin explant workflow (left) with representative images of hematoxylin and eosin staining (right) of freshly collected skin or explants cultured for 48 h. Scale bar, 100 mm. b , Representative immunofluorescence staining of activated Caspase-3 (aCASPASE-3) (red) and DAPI (blue) in freshly collected skin or explants cultured for 48 h showing minimal expression in both conditions. Dotted white line indicates basement membrane. Similar results for donor 1 are not shown. c , Schematic of human skin explant workflow (left), representative images of RNA FISH of CCL19 (red) and PDGFRA (green) on sections from freshly collected skin or explants cultured for 48 h treated with vehicle control or recombinant TNF (10 ng ml −1 ) (center), and quantification of number of PDGFRA + / CCL19 + cells per high-powered field (HPF) (right). Dotted white line indicates basement membrane. Data are mean ± s.e.m. P values shown calculated with two-tailed t -test. d , Additional representative RNA FISH images of superficial perivascular areas of freshly collected skin or human skin explants treated with vehicle control or recombinant TNF (10 ng ml −1 ). e , qPCR of CCL19 in human skin explants cultured in presence of absence of recombinant TNF (10 ng ml −1 ). Data are mean ± s.e.m. *, P < 0.05 by paired one-tailed t -test.

Article Snippet: Blocking was performed with 5.0% normal donkey serum (Jackson ImmunoResearch), 1% BSA (Sigma-Aldrich) and 0.3% Triton X in PBS for 1 h. Sections were incubated overnight at 4 °C with anti-activated Caspase-3 (R&D systems, AF835; 1:100), followed by donkey antirabbit Alexa Fluor 647 secondary antibody (Jackson ImmunoResearch; 1:400) for 1 h at room temperature.

Techniques: Staining, Cell Culture, Immunofluorescence, Expressing, Membrane, Control, Recombinant, Two Tailed Test, One-tailed Test

Journal: Cell Reports Medicine

Article Title: Human liver-derived organoids recapitulate Oropouche virus infection and manifestation, enabling antiviral drug discovery

doi: 10.1016/j.xcrm.2026.102646

Figure Lengend Snippet: Cellular pathogenesis induced by OROV infection in organoids (A) GSEA analysis shows the activation of apoptotic signaling pathways in OROV-infected organoids at 96 h post-inoculation compared to the uninfected controls cultured at the same condition for 96 h. Data are representative of four biological replicates from one RNA-seq experiment. (B) Representative images of organoids 96 h post-inoculation with the OROV-2024 by immunostaining with the antibody against OROV Gc glycoprotein (red), phalloidin for F-actin (green), and DAPI for nuclei (blue). Scale bars, 50 and 10 μm. (C) Representative images of organoids at 1, 6, 12, and 24 h post-inoculation with OROV-2024 by immunostaining with the antibodies against OROV Gc protein (red), cleaved-caspase-3 (green), and DAPI (blue). Scale bars, 50 μm. (D) Representative images of organoids at 48 and 96 h post-inoculation with OROV-2024 by immunostaining with the antibodies against OROV Gc protein (red), cleaved-caspase-3 (green), and DAPI (blue). Scale bars, 25 μm. (E–G) Transmission electron microscopy images of OROV-infected organoids at 96 h post-inoculation, showing damaged mitochondria (M) with disorganized and disintegrating cristae (blue arrows), modified Golgi complexes (G), and extracellular viral particles (red circles). Insets show magnified views. Scale bars: 500 nm and 200 nm (E) and 1 μm and 200 nm (F and G). N, nucleus.

Article Snippet: Primary antibodies used in this study are as follows: Anti-Oropouche orthobunyavirus Gc (1:100, mouse monoclonal antibody; clone 2B5B1; provided by Dr. Sven Reiche from Federal Research Institute for Animal Health, Germany) and Human Active Caspase-3 Antibody (R&D Systems, Bio-Techne, USA).

Techniques: Infection, Activation Assay, Protein-Protein interactions, Cell Culture, RNA Sequencing, Immunostaining, Transmission Assay, Electron Microscopy, Modification

Journal: Cell Reports Medicine

Article Title: Human liver-derived organoids recapitulate Oropouche virus infection and manifestation, enabling antiviral drug discovery

doi: 10.1016/j.xcrm.2026.102646

Figure Lengend Snippet: Molnupiravir attenuated virus-induced cell death and modulates host antiviral responses in OROV-infected liver organoids (A) Representative images of OROV-2024-infected organoids without (top) or with (bottom) 96-h NHC (1 μM) treatment by immunostaining with the antibody against OROV Gc glycoprotein (red), phalloidin for F-actin (green), and DAPI for nuclei (blue). Scale bars: 50 , 25, and 10 μm. (B) Representative images of OROV-2024 infected organoids without (top) or with (bottom) 96-h NHC (1 μM) treatment by immunostaining with the antibody against OROV Gc glycoprotein (red), cleaved-caspase-3 (green), and nuclei (DAPI, blue). Scale bars: 35 and 10 μm. (C) LDH release in culture supernatant of organoids at 48 and 96 h post-inoculation, comparing uninfected, OROV-infected, and NHC-treated groups (1 μM) ( n = 4 biological replicates). (D) GSEA in OROV-infected organoids with NHC treatment for 96 h compared to organoids with OROV infection alone. (E) Heatmap of the top 50 significantly regulated genes comparing uninfected controls, OROV-infected organoids at 96 h post-inoculation, and OROV-infected organoids with 96-h NHC treatment. (F) Heatmap of selected ISGs comparing OROV-infected organoids at 96 h post-inoculation and OROV-infected organoids with 96-h NHC treatment. ISGs were sourced from a published large-scale ISG screen and mapped to our RNA-seq dataset. The color scale represents row-wise Z score of gene expression across samples. Data are representative of four biological replicates from one RNA-seq experiment (D–F). Data represent mean ± SEM; statistical analysis by Mann-Whitney U test; ∗ p < 0.05; ns, not significant.

Article Snippet: Primary antibodies used in this study are as follows: Anti-Oropouche orthobunyavirus Gc (1:100, mouse monoclonal antibody; clone 2B5B1; provided by Dr. Sven Reiche from Federal Research Institute for Animal Health, Germany) and Human Active Caspase-3 Antibody (R&D Systems, Bio-Techne, USA).

Techniques: Virus, Infection, Immunostaining, RNA Sequencing, Gene Expression, MANN-WHITNEY

Journal: Advanced Science

Article Title: Lipid‐Facilitated Opening of the ADAM10 Sheddase Revealed by Enhanced Sampling Simulations

doi: 10.1002/advs.202515713

Figure Lengend Snippet: ADAM10 activity upon apoptosis induction. A) The fluorescence signal of anti‐CD43 DFT1 mAb (CD43, red), anti‐ADAM10 11G2 mAb (ADAM10, blue), PS labelled with annexin‐V (PS, green) and activated caspase‐3 (Casp3, yellow) were recorded 0.5, 1, 2, and 3 h after apoptosis induction using fluorescence confocal microscopy. Scale bar = 7 µm. B) Casp3 vs ADAM10 signal analyzed by flow cytometry after 3 h apoptosis induction. C) Casp3 (orange diamonds), ADAM10 (blue squares) and CD43 (red triangles) signal over time quantified by flow cytometry, expressed as % positive normalized to 100% at T = 0 h. D) Molecular quantification of shed CD43Halo in the supernatant normalized to apoptotic cells using single‐particle profiler of cells in the absence (Non‐Apop) and presence of staurosporine‐induced apoptosis (Apop), with and without the specific ADAM10 inhibitor GI. E) ADAM10 signal quantified by flow cytometry in the absence (NT = non‐treated) or presence of broad‐spectrum ADAM sheddase family inhibitors GM6001 or marimastat in Casp3‐negative (Non‐Apop) and Casp3‐positive (Apoptotic) cells, n = 3–6 independent experiments, expressed as % positive normalized to 100% ADAM10 expression on non‐apoptotic cells. Solid lines represent mean % ADAM10 expression, with significance tested using one‐way ANOVA of log10‐transformed data with Dunnett's post‐hoc test, **** p < 0.0001, ns p > 0.05.

Article Snippet: Fixed cells were washed with PBS, permeabilized with perm buffer (Biolegend 421002), incubated with blocking buffer (5% BSA, 0.015% Triton in PBS) for 45 min and labeled with anti‐human active caspase‐3 antibody clone Asp175 (Cell Signaling Technologies 9661) used at 1:400.

Techniques: Activity Assay, Fluorescence, Confocal Microscopy, Flow Cytometry, Single Particle, Expressing, Transformation Assay