human perforin 1 prf1 elisa kit  (Elabscience Biotechnology)

Journal: Bioactive Materials

Article Title: Dynamic feedback BacGuard anchors microbial metabolism to host symbiosis in real-time ulcerative colitis therapy

doi: 10.1016/j.bioactmat.2026.05.060

Figure Lengend Snippet: BacGuard promotes colon tissue repairment. a) Scheme of microbiota-dependent epithelial repair mechanism orchestrated by BacGuard. b) Short-chain fatty acid (SCFA) profile alterations. n = 6. c) BacGuard-induced probiotic proliferation and d) quantitative results. n = 3. e) Immunofluorescence analysis of ILC3 (ROR γt + CD3 − cells) in colon tissue. f) Flow cytometric analysis of lamina propria lymphocytes (ROR γt + ). n = 3. g) Concentration of IL-22 in MNK-3 cells. n = 3. h) Representative PAS-staining (upper panel) and MUC-2 immunohistochemistry (lower panel) images of colon tissues. n = 5. ns, not significant; ∗p < 0.05, ∗∗p < 0.01 and ∗∗∗p < 0.001.

Article Snippet: For the detection of IL-22 and LPS concentrations, the Mouse IL-22 Precoated ELISA Kit (DAKEWE, China) and Mouse LPS ELISA Kit (JONLNBIO, China) were employed correspondingly.

Techniques: Immunofluorescence, Concentration Assay, Staining, Immunohistochemistry

Journal: iScience

Article Title: Targeting mutant NPM1-expressing AML with T cell receptor-like antibodies

doi: 10.1016/j.isci.2026.117630

Figure Lengend Snippet: Engineering and functional characterization of human-mouse chimeric antibodies (A) Schematic diagram of human-mouse chimeric antibodies. (B) SPR analysis revealed a KD of 492 nM for hu2E2 binding to pNPM1-HLA-A2 (six concentrations were injected once each [ N = 1 per concentration] in a single experiment). (C) Detection of the ADCC effect in Jurkat-Lucia NFAT-CD16 cells (response values were plotted against log10(antibody concentration) and fitted by 4 PL nonlinear regression to illustrate the dose-response relationship. N = 3 independent experiments; data are mean ± SD). (D) The anti-tumor activity of the antibodies in vitro was detected via lymphocyte-tumor coculture. ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; N = 5 independent experiments; data are mean ± SD; ordinary one-way ANOVA. (E) ELISA was used to detect the cytokine content. N = 5 independent experiments; data are represented as mean ± SD.

Article Snippet: Human perforin 1 (PRF1) ELISA kit , Elabscience , Cat# E-EL-H1123.

Techniques: Functional Assay, Analysis, Binding Assay, Injection, Concentration Assay, Activity Assay, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Targeting mutant NPM1-expressing AML with T cell receptor-like antibodies

doi: 10.1016/j.isci.2026.117630

Figure Lengend Snippet: Diverse formats of engineered antibodies (A) Schematic diagram of 2E2-engineered antibodies. (B) Affinity of hu2E2-CAM16vhh to NK cells, as determined via flow cytometry; affinity of hu2E2-H11vhh for T cells, as determined via flow cytometry (one independent experiment; N = 1). (C) Detection of the ADCC effect in Jurkat-Lucia NFAT-CD16 cells ( N = 3 independent experiments; data are mean ± SD). (D) The anti-tumor activity of the antibodies in vitro was detected via lymphocyte-tumor coculture. ∗ p < 0.05; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; N = 5 independent experiments; data are mean ± SD; ordinary one-way ANOVA. (E) ELISA was used to detect the cytokine content. N = 5 independent experiments; data are represented as mean ± SD.

Article Snippet: Human perforin 1 (PRF1) ELISA kit , Elabscience , Cat# E-EL-H1123.

Techniques: Flow Cytometry, Activity Assay, In Vitro, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Targeting mutant NPM1-expressing AML with T cell receptor-like antibodies

doi: 10.1016/j.isci.2026.117630

Figure Lengend Snippet: Effect of Fc region mutation of the 2E2 antibody on the killing of AML cells in vitro (A) Model diagram of the introduction of mutations in the Fc region of the 2E2 antibody. (B) Three Fc-Silence antibodies expressed and purified via the CHO expression system were identified via SDS-PAGE (one independent experiment; N = 1). (C) Detection of the ADCC effect in Jurkat-Lucia NFAT-CD16 cells ( N = 3 independent experiments; data are mean ± SD). (D) The antitumor activity of the antibody in vitro was detected via lymphocyte-tumor coculture. ( N = 5 independent experiments for the hu2E2-CAM16vhh-DANA, hu2E2-H11vhh-DANA, and hu2E2-L1H1-DANA groups, and N = 4 for the Control group. Data are shown as mean ± SD). (E) ELISA was used to measure the levels of cytokines. ∗ p < 0.05;∗∗∗∗ p < 0.0001; N = 5 independent experiments; data are mean ± SD; ordinary one-way ANOVA).

Article Snippet: Human perforin 1 (PRF1) ELISA kit , Elabscience , Cat# E-EL-H1123.

Techniques: Mutagenesis, In Vitro, Purification, Expressing, SDS Page, Activity Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: Targeting mutant NPM1-expressing AML with T cell receptor-like antibodies

doi: 10.1016/j.isci.2026.117630

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Article Snippet: Human perforin 1 (PRF1) ELISA kit , Elabscience , Cat# E-EL-H1123.

Techniques: Cell-Signaling, Purification, Recombinant, Sequencing, Control, Plasmid Preparation, Extraction, Transfection, Bicinchoninic Acid Protein Assay, Conjugation Assay, Staining, Enzyme-linked Immunosorbent Assay, Software

Journal: iScience

Article Title: M2 type tumor-associated macrophage-derived CCL-18 promotes glioma cell proliferation but does not mediates glioma recurrence after radiotherapy

doi: 10.1016/j.isci.2026.117695

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Article Snippet: CCL-18 ELISA kit , Boster Biological Technology , EK0686.

Techniques: Cell-Signaling, Virus, Tissue, Microarray, Recombinant, cDNA Synthesis, Enzyme-linked Immunosorbent Assay, Software, Biomarker Discovery, In Vivo Imaging

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Simultaneous application of IL-4 and Arg promotes efferocytosis and improves the pro-regenerative microenvironment (A) Schematic illustration of the in vitro efferocytosis assessment. (B and C) Macrophage efferocytosis assessment via flow cytometry, showing representative scatterplots of macrophages engulfing apoptotic cells (B) and quantification of the efferocytosis rate after 24 h (C). (D) Morphological evaluation of macrophage efferocytosis, showing representative cell tracking images of macrophages (labeled with CMFDA) engulfing apoptotic cells (labeled with CM-Dil) after 24 h of co-culture. (E) Quantification of the efferocytosis rate after 6, 24, and 48 h determined by flow cytometry under Control and IL-4 + Arg conditions. (F) Experimental design for detecting microenvironmental changes following efferocytosis. (G) ROS quantification following efferocytosis. (H) Enzyme-linked immunosorbent assay (ELISA) quantification of TGF-β1 and IL-10 in the culture supernatants. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (C, G, and H) or two-way ANOVA followed by Sidak’s multiple comparisons test (E). Sample sizes: n = 3 biological replicates (B–E); n = 5 biological replicates, each performed with 3 technical replicates (G) and n = 3 biological replicates, each performed with 3 technical replicates (H). Scale bars, 100 μm (D) and 20 μm (inset).

Article Snippet: Mouse TGF-β1 ELISA Kit , ABclonal , Cat# RK00057.

Techniques: In Vitro, Flow Cytometry, Cell Tracking Assay, Labeling, Co-culture Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Transcriptomic changes underlying synergistic regulation of macrophages by IL-4 and Arg (A) RT-qPCR analysis of M2-related marker gene expression ( Tgfb1 , Il10 , Arg1 , and Cd206 ) in macrophages. (B) Schematic illustration of the bone marrow-derived macrophages (BMDMs) isolation, total RNA extraction, and transcriptomic sequencing workflow. (C) Functional annotation of upregulated genes using Gene Ontology (GO) enrichment analysis. (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis emphasizing significant signaling pathways. (E) RT-qPCR analysis of Tgfb1 and Il10 mRNA expression levels of macrophage after treatment with the IL-4 + Arg combination in the presence or absence of nor-NOHA. (F) ELISA quantification of secreted TGF-β1 and IL-10 protein concentrations in the culture supernatants under nor-NOHA inhibition. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (A, E, and F). Sample sizes: n = 3 biological replicates (C and D); n = 3 (A and F) and 4 (E) biological replicates, each performed with 3 technical replicates.

Article Snippet: Mouse TGF-β1 ELISA Kit , ABclonal , Cat# RK00057.

Techniques: Quantitative RT-PCR, Analysis, Marker, Gene Expression, Derivative Assay, Isolation, RNA Extraction, Sequencing, Functional Assay, Protein-Protein interactions, Expressing, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet:

Article Snippet: Mouse TGF-β1 ELISA Kit , ABclonal , Cat# RK00057.

Techniques: Cell-Signaling, Recombinant, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Software

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: IL-4 and Arg remodel macrophages and promote their migration (A) Schematic illustration of the regulatory role of IL-4 on macrophage phenotype. (B) Volcano plot displaying differentially expressed genes (DEGs) between the control and IL-4-treated macrophages. (C) Heatmap demonstrating the expression of key macrophage polarization markers. (D) Multi-pathway Gene Set Enrichment Analysis (GSEA) demonstrating the integrated enrichment of indicated pathways regulating macrophage functions. (E) Schematic illustration of macrophage priming for efferocytosis mediated by IL-4 and Arg. (F) Relative cell viability of macrophages treated with different concentrations of Arg at 24, 48, and 72 h. (G and H) Macrophage proliferation evaluated by 5-ethynyl-2′-deoxyuridine (EdU) assay, showing representative fluorescence images of EdU and 4',6-diamidino-2-phenylindole (DAPI) staining at 24, 48, and 72 h (G) and quantification of EdU-positive cells treated with 2 mM Arg (H). (I) Schematic illustration of the Transwell migration assay. (J and K) Macrophage recruitment evaluated by Transwell assay, showing representative crystal violet staining images of migrated macrophages after 48 h (J) and quantification of the number of migrated macrophages (K). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (F and H) or one-way ANOVA followed by Tukey’s post hoc test (K). Sample sizes: n = 3 biological replicates (B–D and F); n = 5 biological replicates, each performed with 3 technical replicates (G, H, J, and K). Scale bars, 25 μm (G), 500 μm (J, top), and 100 μm (J, bottom).

Article Snippet: The cumulative release of Arg was quantified using an Arg Content Assay Kit (Solarbio, Beijing, China), and the release of IL-4 was determined using a Mouse IL-4 ELISA Kit (Elabscience, Wuhan, China).

Techniques: Migration, Control, Expressing, Analysis, EdU Assay, Fluorescence, Staining, Transwell Migration Assay, Transwell Assay

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Simultaneous application of IL-4 and Arg promotes efferocytosis and improves the pro-regenerative microenvironment (A) Schematic illustration of the in vitro efferocytosis assessment. (B and C) Macrophage efferocytosis assessment via flow cytometry, showing representative scatterplots of macrophages engulfing apoptotic cells (B) and quantification of the efferocytosis rate after 24 h (C). (D) Morphological evaluation of macrophage efferocytosis, showing representative cell tracking images of macrophages (labeled with CMFDA) engulfing apoptotic cells (labeled with CM-Dil) after 24 h of co-culture. (E) Quantification of the efferocytosis rate after 6, 24, and 48 h determined by flow cytometry under Control and IL-4 + Arg conditions. (F) Experimental design for detecting microenvironmental changes following efferocytosis. (G) ROS quantification following efferocytosis. (H) Enzyme-linked immunosorbent assay (ELISA) quantification of TGF-β1 and IL-10 in the culture supernatants. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (C, G, and H) or two-way ANOVA followed by Sidak’s multiple comparisons test (E). Sample sizes: n = 3 biological replicates (B–E); n = 5 biological replicates, each performed with 3 technical replicates (G) and n = 3 biological replicates, each performed with 3 technical replicates (H). Scale bars, 100 μm (D) and 20 μm (inset).

Article Snippet: The cumulative release of Arg was quantified using an Arg Content Assay Kit (Solarbio, Beijing, China), and the release of IL-4 was determined using a Mouse IL-4 ELISA Kit (Elabscience, Wuhan, China).

Techniques: In Vitro, Flow Cytometry, Cell Tracking Assay, Labeling, Co-culture Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Transcriptomic changes underlying synergistic regulation of macrophages by IL-4 and Arg (A) RT-qPCR analysis of M2-related marker gene expression ( Tgfb1 , Il10 , Arg1 , and Cd206 ) in macrophages. (B) Schematic illustration of the bone marrow-derived macrophages (BMDMs) isolation, total RNA extraction, and transcriptomic sequencing workflow. (C) Functional annotation of upregulated genes using Gene Ontology (GO) enrichment analysis. (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis emphasizing significant signaling pathways. (E) RT-qPCR analysis of Tgfb1 and Il10 mRNA expression levels of macrophage after treatment with the IL-4 + Arg combination in the presence or absence of nor-NOHA. (F) ELISA quantification of secreted TGF-β1 and IL-10 protein concentrations in the culture supernatants under nor-NOHA inhibition. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (A, E, and F). Sample sizes: n = 3 biological replicates (C and D); n = 3 (A and F) and 4 (E) biological replicates, each performed with 3 technical replicates.

Article Snippet: The cumulative release of Arg was quantified using an Arg Content Assay Kit (Solarbio, Beijing, China), and the release of IL-4 was determined using a Mouse IL-4 ELISA Kit (Elabscience, Wuhan, China).

Techniques: Quantitative RT-PCR, Analysis, Marker, Gene Expression, Derivative Assay, Isolation, RNA Extraction, Sequencing, Functional Assay, Protein-Protein interactions, Expressing, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Simultaneous application of IL-4 and Arg promotes bone regeneration in vivo (A) Schematic illustration of the surgical procedure and timeline. (B and C) Evaluation of early macrophage polarization in vivo , showing representative immunofluorescence images of macrophages (F4/80, red) and M2 marker (Arg1, green) at days 1 and 3 post-surgery (B) and quantification of the percentage of M2 macrophages (Arg1 + /F4/80 + ) (C). (D–F) Bone regeneration assessment via micro-CT, showing representative 3D reconstruction images of the bone defect area at day 7 and day 14 post-surgery (D) and quantification of BV (E) and BV/TV (F). (G and H) Histological evaluation of bone repair, showing representative images of H&E and Masson’s trichrome staining at day 7 and day 14 (G) and quantification of new bone area (%) (H). Error bars represent SD. Data were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test (C, E, F, and H). Sample sizes: n = 4 animals per group. Scale bars, 50 μm (B) and 10 μm (inset), 0.5 mm (D) and 200 μm (inset), 250 μm (G).

Article Snippet: The cumulative release of Arg was quantified using an Arg Content Assay Kit (Solarbio, Beijing, China), and the release of IL-4 was determined using a Mouse IL-4 ELISA Kit (Elabscience, Wuhan, China).

Techniques: In Vivo, Immunofluorescence, Marker, Micro-CT, Staining

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet:

Article Snippet: The cumulative release of Arg was quantified using an Arg Content Assay Kit (Solarbio, Beijing, China), and the release of IL-4 was determined using a Mouse IL-4 ELISA Kit (Elabscience, Wuhan, China).

Techniques: Cell-Signaling, Recombinant, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Software

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Simultaneous application of IL-4 and Arg promotes efferocytosis and improves the pro-regenerative microenvironment (A) Schematic illustration of the in vitro efferocytosis assessment. (B and C) Macrophage efferocytosis assessment via flow cytometry, showing representative scatterplots of macrophages engulfing apoptotic cells (B) and quantification of the efferocytosis rate after 24 h (C). (D) Morphological evaluation of macrophage efferocytosis, showing representative cell tracking images of macrophages (labeled with CMFDA) engulfing apoptotic cells (labeled with CM-Dil) after 24 h of co-culture. (E) Quantification of the efferocytosis rate after 6, 24, and 48 h determined by flow cytometry under Control and IL-4 + Arg conditions. (F) Experimental design for detecting microenvironmental changes following efferocytosis. (G) ROS quantification following efferocytosis. (H) Enzyme-linked immunosorbent assay (ELISA) quantification of TGF-β1 and IL-10 in the culture supernatants. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (C, G, and H) or two-way ANOVA followed by Sidak’s multiple comparisons test (E). Sample sizes: n = 3 biological replicates (B–E); n = 5 biological replicates, each performed with 3 technical replicates (G) and n = 3 biological replicates, each performed with 3 technical replicates (H). Scale bars, 100 μm (D) and 20 μm (inset).

Article Snippet: The secretion levels of anti-inflammatory cytokines were quantified using a Mouse TGF-β1 ELISA Kit (ABclonal, Wuhan, China) and a Mouse IL-10 ELISA Kit (Elabscience, Wuhan, China).

Techniques: In Vitro, Flow Cytometry, Cell Tracking Assay, Labeling, Co-culture Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet: Transcriptomic changes underlying synergistic regulation of macrophages by IL-4 and Arg (A) RT-qPCR analysis of M2-related marker gene expression ( Tgfb1 , Il10 , Arg1 , and Cd206 ) in macrophages. (B) Schematic illustration of the bone marrow-derived macrophages (BMDMs) isolation, total RNA extraction, and transcriptomic sequencing workflow. (C) Functional annotation of upregulated genes using Gene Ontology (GO) enrichment analysis. (D) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis emphasizing significant signaling pathways. (E) RT-qPCR analysis of Tgfb1 and Il10 mRNA expression levels of macrophage after treatment with the IL-4 + Arg combination in the presence or absence of nor-NOHA. (F) ELISA quantification of secreted TGF-β1 and IL-10 protein concentrations in the culture supernatants under nor-NOHA inhibition. Error bars represent SD. Data were analyzed by one-way ANOVA followed by Tukey’s post hoc test (A, E, and F). Sample sizes: n = 3 biological replicates (C and D); n = 3 (A and F) and 4 (E) biological replicates, each performed with 3 technical replicates.

Article Snippet: The secretion levels of anti-inflammatory cytokines were quantified using a Mouse TGF-β1 ELISA Kit (ABclonal, Wuhan, China) and a Mouse IL-10 ELISA Kit (Elabscience, Wuhan, China).

Techniques: Quantitative RT-PCR, Analysis, Marker, Gene Expression, Derivative Assay, Isolation, RNA Extraction, Sequencing, Functional Assay, Protein-Protein interactions, Expressing, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: iScience

Article Title: IL-4 and arginine synergy accelerates early bone regeneration via enhanced efferocytosis and ameliorated microenvironment

doi: 10.1016/j.isci.2026.117739

Figure Lengend Snippet:

Article Snippet: The secretion levels of anti-inflammatory cytokines were quantified using a Mouse TGF-β1 ELISA Kit (ABclonal, Wuhan, China) and a Mouse IL-10 ELISA Kit (Elabscience, Wuhan, China).

Techniques: Cell-Signaling, Recombinant, Enzyme-linked Immunosorbent Assay, TUNEL Assay, Software

Journal: iScience

Article Title: Topical application of urolithin A activates mitophagy and enhances mitochondrial function to promote skin longevity

doi: 10.1016/j.isci.2026.117646

Figure Lengend Snippet: UA beneficial impact on human keratinocytes and reconstituted human epidermis (A) Mitochondrial respiration (basal and maximal) assessed as oxygen consumption rate (OCR) in human dermal keratinocytes after 72 h of differentiation from young donor treated with DMSO for 24 h or the indicated dose of UA. ∗ p < 0.05 and ∗∗∗ p < 0.001 after one-way ANOVA. Error bars represent mean ± SEM. (B) TOMM20 immunostaining of skin epidermis in a human full-thickness skin model from an old donor treated with or without UA 1 μM for 25 days. Data expressed as antibody staining signal over imaging field. ∗ p < 0.05 and ∗∗∗∗ p < 0.0001 after unpaired t test one-way ANOVA. Error bars represent mean ± SEM. Scale bars, 50 μm. (C) Relative gene expression of PINK1 and PARK2 in reconstructed human epidermis (RHE). RHE tissues were moisturized for 24 h with or without 1% Mitopure, and relative expression levels of PINK1 and PARK2 were quantified. Statistical analysis was performed using one-way ANOVA (∗ p < 0.05; ∗∗ p < 0.01). Error bars represent mean ± SEM. (D) Interleukin 6 levels normalized to cell viability in keratinocytes. Keratinocytes were stimulated with a pro-inflammatory cytokine mix for 24 h then treated for an additional 24 h with either DMSO or 1 μM UA. IL-6 secretion was quantified and normalized to cell viability. Statistical analysis was performed using one-way ANOVA (∗∗∗∗ p < 0.0001). Error bars represent mean ± SEM. (E) p16 immunostaining in a human full-thickness skin model from an old donor treated with or without UA 1 μM for 25 days. Data expressed as antibody staining signal over imaging field. ∗∗∗∗ p < 0.0001 after unpaired t test. Error bars represent mean ± SEM. Scale bars, 50 μm. (F) Ki67 immunostaining in a human full-thickness skin model from an old donor treated with or without UA 1 μM for 25 days. Data expressed as number of Ki67 positive cells normalized over μm of dermal epidermal junction (count/μm) ∗∗∗∗ p < 0.0001 after unpaired t test. Error bars represent mean ± SEM. Scale bars, 50 μm. (G) Keratin 10 immunostaining in a human full-thickness skin model from an old donor treated with or without UA 1 μM for 25 days. Data expressed as fluorescence intensity over epidermis area. ∗∗∗∗ p < 0.0001 after unpaired t test. Error bars represent mean ± SEM. Scale bars, 50 μm.

Article Snippet: IL-6 ELISA Kit , Proteintech , Cat#KE10007.

Techniques: Immunostaining, Staining, Imaging, Gene Expression, Expressing, Analysis, Fluorescence

Journal: iScience

Article Title: Topical application of urolithin A activates mitophagy and enhances mitochondrial function to promote skin longevity

doi: 10.1016/j.isci.2026.117646

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Article Snippet: IL-6 ELISA Kit , Proteintech , Cat#KE10007.

Techniques: Binding Assay, Ex Vivo, Recombinant, Metals, Enzyme-linked Immunosorbent Assay, Reverse Transcription, Cell Viability Assay, Staining, DC Protein Assay, Gene Expression, Glass, Software, Imaging, Microscopy, Real-time Polymerase Chain Reaction

Journal: MedComm

Article Title: Blocking Extracellular Cyclophilin A‐Mediated Adhesion Protects Against Bacterial Infection

doi: 10.1002/mco2.71063

Figure Lengend Snippet: eCypA is critical for the adhesion of GAS and S.P (A) C57BL/6 mice ( n = 7) were infected intranasally with PBS or 3000 PFU of influenza A/WSN/33 (H1N1) on Day 0 followed by intraperitoneal injection of isotype IgG (IgG) or anti‐CypA mAb (100 µg/mouse) on Day 1, and then infected intranasally with GAS (1.0 × 10 7 CFU), S. P (1.0 × 10 7 CFU), K. P (1.0 × 10 7 CFU), or PBS on Day 3. (B) Adherence of GAS in the lungs of C57BL/6 mice on Day 4. (C) Adherence of S. P in the lungs of C57BL/6 mice on Day 4. (D) The lung index (100× lung/body weight) was calculated on Day 4. (E) The body weights of C57BL/6 mice were monitored during Days 0–4. (F) Hematoxylin and eosin staining of mouse lungs on Day 4 (left). The severity of lung injury was analyzed in a blinded manner (right). Scale bar, 150 µm. (G) Heatmap showing the results of the qPCR analysis indicating the mRNA levels of proinflammatory cytokines in the lungs of C57BL/6 mice on Day 4. (H and I) Adherence of GAS (H) and S. P (I) in A549 cells infected with PBS or IAV (MOI = 1) followed by treatment with IgG or anti‐CypA mAb (50 nM) for 24 h and then infection of the cells with GAS (MOI = 10) for 2 h. Data are reported as the percentage of adherent bacteria relative to the input inoculum. (J) C57BL/6 mice were preinjected intraperitoneally with eCypA (40 µg/mouse) or vehicle (PBS) for 2 h, followed by intraperitoneal injection with anti‐CypA mAb (100 µg/mouse) or vehicle (PBS) on Day 0 and then infected intranasally with GAS (1.0 × 10 7 CFU), S. P (1.0 × 10 7 CFU), K. P (1.0 × 10 7 CFU), or PBS on Day 3. (K) ELISA of eCypA in BALF from mice infected with GAS. (L–N) Adherence of GAS (K), S. P (L), and K. P (M) in the lungs of C57BL/6 mice ( n = 5) on Day 4. (O–Q) Adherence assays of GAS (N), S. P (O), and K. P (P) in A549 cells pretreated with PBS, eCypA (10 nM) or eCypA + anti‐CypA mAb (10 nM eCypA + 50 nM anti‐CypA mAb, preincubated at 37°C for 1 h) for 12 h, followed by GAS, S. P , or K. P (MOI = 10) infection for 2 h. Data are shown as the percentage of adherent bacteria relative to the input inoculum. Data shown in (B–F) and (K–N) are presented as the means ± SEMs; Data shown in (H), (I), and (O–Q) are presented as the means ± SDs; * p < 0.05, ** p < 0.01, *** p < 0.001 and ns = not significant. Unpaired two‐tailed Student's t test for (B) and (C), and one‐way ANOVA followed by Dunnett's test for (D–F), (H), (I), and (K–Q).

Article Snippet: eCypA concentrations in BALF, serum, and cell culture supernatants were quantified by ELISA (CSB‐E09920h, CSB‐E09282m, CUSABIO ).

Techniques: Infection, Injection, H&E Stain, Analysis, Bacteria, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: MedComm

Article Title: Blocking Extracellular Cyclophilin A‐Mediated Adhesion Protects Against Bacterial Infection

doi: 10.1002/mco2.71063

Figure Lengend Snippet: Binding of eCypA to TβR2 facilitates the formation of the TGFβ1‐TGF‐β receptor 2 complex (A) qPCR analysis of ITGA5 and FN mRNA levels in A549 cells transfected with siRNAs targeting TβR2, CD147, ACE2, or MUC18 for 48 h, and then treated with 10 nM eCypA or PBS for 12 h. (B) Immunoblot analysis of the indicated proteins in immunoprecipitated samples and whole‐cell lysates (WCL) of A549 cells treated with Fc, CypA‐Fc (200 ng/mL), R55‐Fc (200 ng/mL), CypA‐Fc + anti‐CypA mAb (1 µg/mL), or R55‐Fc + anti‐CypA mAb for 2 h. (C) Interactions of CypA with TGF‐β1, TβR1, and TβR2 were detected using ELISA. BSA was used as a negative control. (D) The blocking effect of the anti‐CypA mAb on the binding of CypA to TβR2 was detected using ELISA. (E–H) Interaction of CypA with anti‐CypA mAb (E) and TβR2 (F) was detected by SPR. The competitive effect of anti‐CypA mAb on CypA‐TβR2 interaction was detected using competitive SPR assay (G). Linearities of the CypA‐TβR2 interaction were generated from the data of the sampling points shown in (H). (I) Immunoblot analysis of the indicated proteins in A549 cells treated with eCypA (10 nM) or eCypA (10 nM) + TGF‐β pAb (50 nM) for 30 min. (J) Adherence assays of GAS in A549 cells treated with eCypA (10 nM) or eCypA (10 nM) + TGF‐β pAb (50 nM) for 12 h, and then infected with GAS (MOI = 10.0) for 2 h. Data are reported as the percentage of adherent bacteria relative to the input inoculum. (K) Immunoblot analysis of the indicated proteins in immunoprecipitated samples and WCLs of A549 cells treated with TGF‐β1 (5 ng/mL), TGF‐β1 + CypA‐His (10 nM), or TGF‐β1 + CypA‐His + anti‐CypA mAb (50 nM) for 2 h. (L and M) Interactions of TGF‐β1 with TβR2 were detected by SPR in the absence (L) or presence (M) of CypA. Data are presented as the means ± SDs; ** p < 0.01, *** p < 0.001 and ns = not significant; unpaired two‐tailed Student's t test for (D); one‐way ANOVA followed by Dunnett's test for (A) and (J).

Article Snippet: eCypA concentrations in BALF, serum, and cell culture supernatants were quantified by ELISA (CSB‐E09920h, CSB‐E09282m, CUSABIO ).

Techniques: Binding Assay, Analysis, Transfection, Western Blot, Immunoprecipitation, Enzyme-linked Immunosorbent Assay, Negative Control, Blocking Assay, SPR Assay, Generated, Sampling, Infection, Bacteria, Two Tailed Test