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human embryonic kidney hek293 cells  (ATCC)


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    Structured Review

    ATCC human embryonic kidney hek293 cells
    MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to <t>HEK293</t> culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = <t>human</t> <t>embryonic</t> <t>kidney;</t> miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.
    Human Embryonic Kidney Hek293 Cells, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1852 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle"

    Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

    Journal: Journal of Sport and Health Science

    doi: 10.1016/j.jshs.2025.101091

    MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to HEK293 culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = human embryonic kidney; miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.
    Figure Legend Snippet: MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to HEK293 culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = human embryonic kidney; miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.

    Techniques Used: Cell Culture, Fluorescence, Derivative Assay, Labeling, Control, Staining

    NRDC is a direct target of miR-136-3p in human myotubes. Skeletal muscle NRDC mRNA is responsive to training and inactivity. (A) Tissue mRNA expression of NRDC from the Human Protein Atlas database showing enriched expression of NRDC in human skeletal muscle. (B) The miR-136-3p target site in the NRDC gene is highly conserved in mammals. (C) Luciferase activity in HEK293 cells co-transfected the NRDC 3’UTR and miR-136-3p with or without anti-miR136-3p inhibitors. miR-136-3p transfection downregulates NRDC (D) mRNA and (E) representative image of protein abundance in human myotubes. (F) Publicly available data ( GSE14413 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 6 weeks of endurance training ( n = 8). (G) Publicly available data ( GSE120862 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 2 months of aerobic training ( n = 10). (H) Publicly available data ( GSE14901 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 14 days of immobilization ( n = 24). * p < 0.05, ** p < 0.005. GSE = gene set enrichment; HEK293 = human embryonic kidney; miR = microRNA; NC = negative control; NRDC = nardilysin convertase; nTPM = normalized transcripts per million; si NRDC = small interfering RNA of NRDC ; UTR = untranslated region.
    Figure Legend Snippet: NRDC is a direct target of miR-136-3p in human myotubes. Skeletal muscle NRDC mRNA is responsive to training and inactivity. (A) Tissue mRNA expression of NRDC from the Human Protein Atlas database showing enriched expression of NRDC in human skeletal muscle. (B) The miR-136-3p target site in the NRDC gene is highly conserved in mammals. (C) Luciferase activity in HEK293 cells co-transfected the NRDC 3’UTR and miR-136-3p with or without anti-miR136-3p inhibitors. miR-136-3p transfection downregulates NRDC (D) mRNA and (E) representative image of protein abundance in human myotubes. (F) Publicly available data ( GSE14413 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 6 weeks of endurance training ( n = 8). (G) Publicly available data ( GSE120862 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 2 months of aerobic training ( n = 10). (H) Publicly available data ( GSE14901 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 14 days of immobilization ( n = 24). * p < 0.05, ** p < 0.005. GSE = gene set enrichment; HEK293 = human embryonic kidney; miR = microRNA; NC = negative control; NRDC = nardilysin convertase; nTPM = normalized transcripts per million; si NRDC = small interfering RNA of NRDC ; UTR = untranslated region.

    Techniques Used: Expressing, Luciferase, Activity Assay, Transfection, Quantitative Proteomics, Negative Control, Small Interfering RNA



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    ATCC human embryonic kidney hek293 cells
    MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to <t>HEK293</t> culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = <t>human</t> <t>embryonic</t> <t>kidney;</t> miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.
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    iec  (ATCC)
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    MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to <t>HEK293</t> culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = <t>human</t> <t>embryonic</t> <t>kidney;</t> miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.
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    ATCC murine pre osteoblasts
    Curvature-controlled orientation of cytoskeletal stress fibers on concave-cylindrical surfaces. (a) Representative confocal microscopy images depicting F-actin (magenta) and nuclei (blue) of fibroblasts, mesenchymal stromal cells, <t>osteoblasts,</t> <t>pre-osteoblasts</t> and endothelial cells seeded on flat surfaces. (b) Brass mold used to fabricate the master GeoChip from which GeoChips for use in cell culture are manufactured via sugar candy molding . Photographs show the topographic surface of the brass mold and the candy mold (Scale bar: 2 mm). Scanning electron microscopy (SEM) verified the smoothness of the resulting curved surface (half-cylinder with Ø = 1000 μm, scale bar: 200 μm). (c) Representative confocal microscopy images of cells seeded on concave-cylindrical surfaces with Ø = 100 and 1000 μm. Yellow dashed lines indicate the half-cylinder boundaries. (d-i) Distribution of stress fiber orientation quantified from the F-actin signal of cells on substrates with increasing curvature (average with standard deviation). Cartesian plots include data for fibroblasts (blue), mesenchymal stromal cells (green), osteoblasts (purple), pre-osteoblasts (orange) and endothelial cells (red). The direction 0° - 180° represents the orientation along the cylindrical surface (minimum curvature) and the direction 90° represents the orientation perpendicular to the cylindrical surface (maximum curvature). The substrate curvature experienced in dependency of the orientation is indicated by the red dashed line and red scale. Random orientation is indicated by the black dashed line. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/cell type, 1 donor/cell type. Scale bars 100 μm (unless otherwise stated).
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    ATCC nih 3t3 fibroblasts
    In vitro cell evaluations. (a, b) Fluorescence microscopic images of <t>NIH</t> <t>3T3</t> cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.
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    nih3t3  (ATCC)
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    Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a <t>NIH3T3</t> cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.
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    htr  (ATCC)
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    CBNK-EVs rapidly impair ZIKV infectivity and protect host cells. (a) Multiplex immunohistochemical analysis of ZIKV infection in neural progenitor cells. Co-staining for Nestin (neural progenitor cells, green) and ZIKV E protein (red) was performed, and double-positive cells were quantified across six random fields, mean ± SD, ∗P < 0.05. Scale bar, 20 μm. (b) KEGG pathway enrichment bubble plot from <t>RNA-seq</t> <t>of</t> <t>HTR-8/Svneo</t> cells treated with CBNK-EVs (3 × 10 11 particles/mL, 10 μl) and ZIKV (MOI = 1) for 1 h. (c) GSEA analysis of transcriptomic data from HTR-8/Svneo cells. (d) Western blot analysis of ZIKV E and ZIKV NS5 protein levels after co-incubation of ZIKV particles with CBNK-EVs for the indicated time periods (0–30 min) prior to infection. (e) The abundance of NK cell-associated proteins in CBNK-EVs. ∗P < 0.05 (one-way ANOVA).
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    CBNK-EVs rapidly impair ZIKV infectivity and protect host cells. (a) Multiplex immunohistochemical analysis of ZIKV infection in neural progenitor cells. Co-staining for Nestin (neural progenitor cells, green) and ZIKV E protein (red) was performed, and double-positive cells were quantified across six random fields, mean ± SD, ∗P < 0.05. Scale bar, 20 μm. (b) KEGG pathway enrichment bubble plot from <t>RNA-seq</t> <t>of</t> <t>HTR-8/Svneo</t> cells treated with CBNK-EVs (3 × 10 11 particles/mL, 10 μl) and ZIKV (MOI = 1) for 1 h. (c) GSEA analysis of transcriptomic data from HTR-8/Svneo cells. (d) Western blot analysis of ZIKV E and ZIKV NS5 protein levels after co-incubation of ZIKV particles with CBNK-EVs for the indicated time periods (0–30 min) prior to infection. (e) The abundance of NK cell-associated proteins in CBNK-EVs. ∗P < 0.05 (one-way ANOVA).
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    293t  (ATCC)
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    Perforin mediates the direct virion-disrupting activity of CBNK-EVs. (a) Quantification of perforin and granzyme B in CBNK-EVs by ELISA. (b) Vero-E6 cells were infected with ZIKV (MOI = 1) after pre-incubation of viral particles with 40 ng/L of recombinant perforin or granzyme B for 2 h. ZIKV E protein levels were assessed by Western blot after 24 h, densitometric analysis of the protein bands is shown (right, n = 3). (c) CBNK-EVs were pre-incubated with ZIKV in the presence or absence of 10 mM EGTA, followed by micrococcal nuclease digestion. Protected ZIKV E RNA was quantified by RT-qPCR to assess virion integrity. (d) Western blot analysis of perforin expression in CBNK cells after transfection with the indicated siRNAs. (e) Quantification of perforin levels from (d). (f) Perforin levels in CBNK-EVs collected from control or perforin-knockdown cells, measured by ELISA and normalized to particle count (per 10 11 particles, n = 6). (g) Vero-E6 cells were infected with ZIKV (MOI = 1) that had been pre-incubated with control or perforin-knockdown CBNK-EVs. ZIKV E protein levels were evaluated by Western blot. (h) Analysis of ZIKV E protein levels from (g). (i) Characterization of ITGB2-EVs and control EVs from <t>293T</t> cells by NTA and TEM. Scale bar, 100 nm. (j) Zeta potential measurements of ITGB2-EVs and control EVs. (k) Western blot analysis of EV markers and ITGB2 expression in ITGB2-EVs and control EVs. (l) Antiviral activity of CBNK-EVs and ITGB2-EVs evaluated by cell-based ZIKV E protein ELISA. Data are presented as mean ± SD (n = 3). ∗P < 0.05, ∗∗P < 0.01, ∗P < 0.001 (one-way ANOVA).
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    ATCC human b lymphoblastoid cell line tk6
    Micronucleus test of Kratom leaf extract after 4 h exposure with S9 in <t>TK6</t> cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).
    Human B Lymphoblastoid Cell Line Tk6, supplied by ATCC, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC vero e6 cells
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    Image Search Results


    MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to HEK293 culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = human embryonic kidney; miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.

    Journal: Journal of Sport and Health Science

    Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

    doi: 10.1016/j.jshs.2025.101091

    Figure Lengend Snippet: MiR-136-3p content in cell culture medium and uptake of extracellular miR-136-3p into cultured myotubes. MiRNA content in (A) human myotubes and (B) human pancreatic islets culture media. Results were first normalized using RNU1A1 and then presented in relation to miR-23a-3p content for n = 4 different donors for myotubes cultures and n = 4 donors for human islets. Left panel (C) shows bright-field image of cultured human myotubes and right panel (C) shows a representative fluorescence image of cultured human myotubes with cells exposed to human serum-derived EVs loaded with Cy3-miR-136-3p. Cy3 fluorescence (red) is detected in the whole cytoplasm of the human myotubes. (D) Representative fluorescence image of human myotubes exposed to HEK293 culture medium with EVs loaded with Cy3-miR-136-3p (red). (E) Representative image of human myotubes exposed to EVs loaded with TexasRed-labeled with a control RNA (orange). Nuclear Hoechst staining is shown in blue. Scale bar = 100 μm. EVs = extracellular vesicles; HEK293 = human embryonic kidney; miR = microRNA; Rel = relative; RNU1A1 = U1 small nuclear RNA.

    Article Snippet: Human embryonic kidney (HEK293) cells were obtained from American Type Culture Collection (ATCC) and cultured in high-glucose (4.5 g/L) Dulbecco's Modified Eagle Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% (vol/vol) FBS.

    Techniques: Cell Culture, Fluorescence, Derivative Assay, Labeling, Control, Staining

    NRDC is a direct target of miR-136-3p in human myotubes. Skeletal muscle NRDC mRNA is responsive to training and inactivity. (A) Tissue mRNA expression of NRDC from the Human Protein Atlas database showing enriched expression of NRDC in human skeletal muscle. (B) The miR-136-3p target site in the NRDC gene is highly conserved in mammals. (C) Luciferase activity in HEK293 cells co-transfected the NRDC 3’UTR and miR-136-3p with or without anti-miR136-3p inhibitors. miR-136-3p transfection downregulates NRDC (D) mRNA and (E) representative image of protein abundance in human myotubes. (F) Publicly available data ( GSE14413 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 6 weeks of endurance training ( n = 8). (G) Publicly available data ( GSE120862 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 2 months of aerobic training ( n = 10). (H) Publicly available data ( GSE14901 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 14 days of immobilization ( n = 24). * p < 0.05, ** p < 0.005. GSE = gene set enrichment; HEK293 = human embryonic kidney; miR = microRNA; NC = negative control; NRDC = nardilysin convertase; nTPM = normalized transcripts per million; si NRDC = small interfering RNA of NRDC ; UTR = untranslated region.

    Journal: Journal of Sport and Health Science

    Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

    doi: 10.1016/j.jshs.2025.101091

    Figure Lengend Snippet: NRDC is a direct target of miR-136-3p in human myotubes. Skeletal muscle NRDC mRNA is responsive to training and inactivity. (A) Tissue mRNA expression of NRDC from the Human Protein Atlas database showing enriched expression of NRDC in human skeletal muscle. (B) The miR-136-3p target site in the NRDC gene is highly conserved in mammals. (C) Luciferase activity in HEK293 cells co-transfected the NRDC 3’UTR and miR-136-3p with or without anti-miR136-3p inhibitors. miR-136-3p transfection downregulates NRDC (D) mRNA and (E) representative image of protein abundance in human myotubes. (F) Publicly available data ( GSE14413 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 6 weeks of endurance training ( n = 8). (G) Publicly available data ( GSE120862 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 2 months of aerobic training ( n = 10). (H) Publicly available data ( GSE14901 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 14 days of immobilization ( n = 24). * p < 0.05, ** p < 0.005. GSE = gene set enrichment; HEK293 = human embryonic kidney; miR = microRNA; NC = negative control; NRDC = nardilysin convertase; nTPM = normalized transcripts per million; si NRDC = small interfering RNA of NRDC ; UTR = untranslated region.

    Article Snippet: Human embryonic kidney (HEK293) cells were obtained from American Type Culture Collection (ATCC) and cultured in high-glucose (4.5 g/L) Dulbecco's Modified Eagle Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% (vol/vol) FBS.

    Techniques: Expressing, Luciferase, Activity Assay, Transfection, Quantitative Proteomics, Negative Control, Small Interfering RNA

    Curvature-controlled orientation of cytoskeletal stress fibers on concave-cylindrical surfaces. (a) Representative confocal microscopy images depicting F-actin (magenta) and nuclei (blue) of fibroblasts, mesenchymal stromal cells, osteoblasts, pre-osteoblasts and endothelial cells seeded on flat surfaces. (b) Brass mold used to fabricate the master GeoChip from which GeoChips for use in cell culture are manufactured via sugar candy molding . Photographs show the topographic surface of the brass mold and the candy mold (Scale bar: 2 mm). Scanning electron microscopy (SEM) verified the smoothness of the resulting curved surface (half-cylinder with Ø = 1000 μm, scale bar: 200 μm). (c) Representative confocal microscopy images of cells seeded on concave-cylindrical surfaces with Ø = 100 and 1000 μm. Yellow dashed lines indicate the half-cylinder boundaries. (d-i) Distribution of stress fiber orientation quantified from the F-actin signal of cells on substrates with increasing curvature (average with standard deviation). Cartesian plots include data for fibroblasts (blue), mesenchymal stromal cells (green), osteoblasts (purple), pre-osteoblasts (orange) and endothelial cells (red). The direction 0° - 180° represents the orientation along the cylindrical surface (minimum curvature) and the direction 90° represents the orientation perpendicular to the cylindrical surface (maximum curvature). The substrate curvature experienced in dependency of the orientation is indicated by the red dashed line and red scale. Random orientation is indicated by the black dashed line. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/cell type, 1 donor/cell type. Scale bars 100 μm (unless otherwise stated).

    Journal: Bioactive Materials

    Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

    doi: 10.1016/j.bioactmat.2026.02.005

    Figure Lengend Snippet: Curvature-controlled orientation of cytoskeletal stress fibers on concave-cylindrical surfaces. (a) Representative confocal microscopy images depicting F-actin (magenta) and nuclei (blue) of fibroblasts, mesenchymal stromal cells, osteoblasts, pre-osteoblasts and endothelial cells seeded on flat surfaces. (b) Brass mold used to fabricate the master GeoChip from which GeoChips for use in cell culture are manufactured via sugar candy molding . Photographs show the topographic surface of the brass mold and the candy mold (Scale bar: 2 mm). Scanning electron microscopy (SEM) verified the smoothness of the resulting curved surface (half-cylinder with Ø = 1000 μm, scale bar: 200 μm). (c) Representative confocal microscopy images of cells seeded on concave-cylindrical surfaces with Ø = 100 and 1000 μm. Yellow dashed lines indicate the half-cylinder boundaries. (d-i) Distribution of stress fiber orientation quantified from the F-actin signal of cells on substrates with increasing curvature (average with standard deviation). Cartesian plots include data for fibroblasts (blue), mesenchymal stromal cells (green), osteoblasts (purple), pre-osteoblasts (orange) and endothelial cells (red). The direction 0° - 180° represents the orientation along the cylindrical surface (minimum curvature) and the direction 90° represents the orientation perpendicular to the cylindrical surface (maximum curvature). The substrate curvature experienced in dependency of the orientation is indicated by the red dashed line and red scale. Random orientation is indicated by the black dashed line. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/cell type, 1 donor/cell type. Scale bars 100 μm (unless otherwise stated).

    Article Snippet: Murine pre-osteoblasts (MC3T3-E1, CRL-2593TM, ATCC) were cultured in alpha modified minimum essential medium with nucleosides (F 0925, Biochrom AG), supplemented with 10 % v/v FBS, 1 % v/v P/S and 1 % v/v GlutaMAX (35050, Gibco®).

    Techniques: Confocal Microscopy, Cell Culture, Electron Microscopy, Standard Deviation, MANN-WHITNEY

    Incidence of cell spanning on concave-cylindrical surfaces. (a) Lateral view of fibroblasts exposed to cylinders with increasing diameter (decreasing curvature), with spanning cells marked by yellow arrows. (b) Probability of spanning cells in relation to the half-cylinder diameter. (c-e, top to bottom) Representative 3D reconstructed images of fibroblasts, pre-osteoblasts and endothelial cells on concave-cylindrical surfaces with Ø = 100, 200 and 300 μm. Cells were reconstructed in Imaris using the F-actin (magenta, cell surface reconstruction) and nuclei (blue) signal as obtained by confocal microscopy. Half-cylinder contour is indicated by the yellow dashed line. Spanning cells are indicated by yellow arrows in subfigures c-e for clarity. Polar plots on the right depict the percentage of spanning cells and the corresponding angle of cell orientation for fibroblasts (blue), pre-osteoblasts (orange) and endothelial cells (red). The direction 0° - 180° represents the orientation along the cylindrical surface (minimum curvature) and the direction −90° - 90° represents the orientation perpendicular to the cylindrical surface (maximum curvature). (f) Confocal microscopy images of representative cell morphologies for fibroblasts, pre-osteoblasts and endothelial cells depicting F-actin (magenta), nuclei (blue) and focal adhesions via vinculin staining (green). Focal adhesions are indicated by green arrows (example shown on fibroblasts). (g) Cell length quantified as the major axis of an ellipse fitted around the cell. (h) Cell roundness with a value of 1 representing a perfect circle and value of 0 representing a straight line. (i) FSD calculated as the distance between FA clusters (see methods part for detailed description). (j) FA size distribution per cell plotted as the percentage of FAs that fall into the indicated size classes. (k) Representative force vector maps and (l) total cell force quantified via TFM. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/cell type. N ≥ 60 cells/cell type for FA and morphological analysis. 1 donor/cell type. Scale bar 50 μm.

    Journal: Bioactive Materials

    Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

    doi: 10.1016/j.bioactmat.2026.02.005

    Figure Lengend Snippet: Incidence of cell spanning on concave-cylindrical surfaces. (a) Lateral view of fibroblasts exposed to cylinders with increasing diameter (decreasing curvature), with spanning cells marked by yellow arrows. (b) Probability of spanning cells in relation to the half-cylinder diameter. (c-e, top to bottom) Representative 3D reconstructed images of fibroblasts, pre-osteoblasts and endothelial cells on concave-cylindrical surfaces with Ø = 100, 200 and 300 μm. Cells were reconstructed in Imaris using the F-actin (magenta, cell surface reconstruction) and nuclei (blue) signal as obtained by confocal microscopy. Half-cylinder contour is indicated by the yellow dashed line. Spanning cells are indicated by yellow arrows in subfigures c-e for clarity. Polar plots on the right depict the percentage of spanning cells and the corresponding angle of cell orientation for fibroblasts (blue), pre-osteoblasts (orange) and endothelial cells (red). The direction 0° - 180° represents the orientation along the cylindrical surface (minimum curvature) and the direction −90° - 90° represents the orientation perpendicular to the cylindrical surface (maximum curvature). (f) Confocal microscopy images of representative cell morphologies for fibroblasts, pre-osteoblasts and endothelial cells depicting F-actin (magenta), nuclei (blue) and focal adhesions via vinculin staining (green). Focal adhesions are indicated by green arrows (example shown on fibroblasts). (g) Cell length quantified as the major axis of an ellipse fitted around the cell. (h) Cell roundness with a value of 1 representing a perfect circle and value of 0 representing a straight line. (i) FSD calculated as the distance between FA clusters (see methods part for detailed description). (j) FA size distribution per cell plotted as the percentage of FAs that fall into the indicated size classes. (k) Representative force vector maps and (l) total cell force quantified via TFM. Statistical significance via Mann-Whitney test (two sided) with Bonferroni correction, ∗p < 0.05. N ≥ 3 GeoChips/cell type for a total of N ≥ 12 half-cylinders/cell type. N ≥ 60 cells/cell type for FA and morphological analysis. 1 donor/cell type. Scale bar 50 μm.

    Article Snippet: Murine pre-osteoblasts (MC3T3-E1, CRL-2593TM, ATCC) were cultured in alpha modified minimum essential medium with nucleosides (F 0925, Biochrom AG), supplemented with 10 % v/v FBS, 1 % v/v P/S and 1 % v/v GlutaMAX (35050, Gibco®).

    Techniques: Confocal Microscopy, Staining, Plasmid Preparation, MANN-WHITNEY

    Cell spanning initiates channel closure and subsequent tissue remodeling. (a) Fabrication of full-cylindrical channels with Ø = 250 μm in PDMS substrates by direct molding from a micro-machined brass mold. (b) Degree of channel closure representing the distribution of cells within the channels at the selected points in time during live confocal imaging. A value of 0 indicates that cells are exclusively found at the wall of the channel and a value of 1 indicates cells have completely closed the channel and are homogeneously distributed. (c) Relative degree of alignment of the cell-network within the channels quantified as the maximum value of the orientation distribution for the individual cell types and time points normalized to the highest detected value of all conditions (see also Supplementary Data S2). Higher values indicate a higher degree of alignment along the channel axis. (d-f) Lateral and front view of the PDMS cylindrical channels obtained by live confocal imaging of fibroblasts (blue), pre-osteoblasts (orange) and endothelial cells (red) using CellTracker™ Green ( t = 4, 12, 24 and 48 h after seeding). Open arrows indicate cells spanning perpendicular to the channel axis. Full arrows indicate cells oriented along the direction of the channel axis after channel closure. Channel contour is highlighted by the yellow dashed lines. The surface of the forming tissue is marked by red dashed lines. White dashed lines indicate the z-volume that is shown in the corresponding lateral views. Statistical significance via Mann-Whitney test with Bonferroni correction, ∗p < 0.05. N = 3 cylindrical channels/cell type. 1 donor/cell type. Scale bars 100 μm.

    Journal: Bioactive Materials

    Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

    doi: 10.1016/j.bioactmat.2026.02.005

    Figure Lengend Snippet: Cell spanning initiates channel closure and subsequent tissue remodeling. (a) Fabrication of full-cylindrical channels with Ø = 250 μm in PDMS substrates by direct molding from a micro-machined brass mold. (b) Degree of channel closure representing the distribution of cells within the channels at the selected points in time during live confocal imaging. A value of 0 indicates that cells are exclusively found at the wall of the channel and a value of 1 indicates cells have completely closed the channel and are homogeneously distributed. (c) Relative degree of alignment of the cell-network within the channels quantified as the maximum value of the orientation distribution for the individual cell types and time points normalized to the highest detected value of all conditions (see also Supplementary Data S2). Higher values indicate a higher degree of alignment along the channel axis. (d-f) Lateral and front view of the PDMS cylindrical channels obtained by live confocal imaging of fibroblasts (blue), pre-osteoblasts (orange) and endothelial cells (red) using CellTracker™ Green ( t = 4, 12, 24 and 48 h after seeding). Open arrows indicate cells spanning perpendicular to the channel axis. Full arrows indicate cells oriented along the direction of the channel axis after channel closure. Channel contour is highlighted by the yellow dashed lines. The surface of the forming tissue is marked by red dashed lines. White dashed lines indicate the z-volume that is shown in the corresponding lateral views. Statistical significance via Mann-Whitney test with Bonferroni correction, ∗p < 0.05. N = 3 cylindrical channels/cell type. 1 donor/cell type. Scale bars 100 μm.

    Article Snippet: Murine pre-osteoblasts (MC3T3-E1, CRL-2593TM, ATCC) were cultured in alpha modified minimum essential medium with nucleosides (F 0925, Biochrom AG), supplemented with 10 % v/v FBS, 1 % v/v P/S and 1 % v/v GlutaMAX (35050, Gibco®).

    Techniques: Imaging, MANN-WHITNEY

    Channel closure mechanism can be controlled by substrate curvature using scaffolds with well-defined geometries. (a, left) Schematic representation of the in vitro culture setup with collagen scaffold presenting channels of controlled diameter with Ø ≈ 600 μm, Ø ≈ 350 μm and Ø ≈ 150 μm. Monolayer seeding on one side of the biomaterial facilitates migration of cells from one end of the biomaterial. (a, right) SEM image of the microarchitecture (Scale bar 20 μm) and channels within the biomaterial (Scale bars 100 μm). SEM images correspond to the outermost surface of the scaffold. (b) Comparison of template diameter against resulting channel diameter after cross-linking and sterilization of the biomaterial. (c) Representative images of fibroblasts, pre-osteoblasts and endothelial cells within channels of distinct diameters 7 days after seeding. Cell cytoskeleton (F-actin) is depicted in magenta and nuclei in blue. Yellow arrows indicate the direction (arrow angle) and degree of alignment (vector length) for the corresponding region. Scale bar close-up images: 25 μm. (d, left) Degree of channel closure for the investigated channel diameters and cell types. (d, right) Relative degree of tissue alignment for the different channel diameters and cell types. Tissue alignment ranges from 0 (fully isotropic) to 1 (fully anisotropic, dashed line). Tissue across the channel and relative degree of is calculated in the central 50 % of each channel. Data displayed as average with standard deviation. N = 4 scaffolds/cell type. 1 donor/cell type. Scale bars 200 μm (unless otherwise stated).

    Journal: Bioactive Materials

    Article Title: Cell type-specific response to curvature controls tissue growth dynamics in biomaterial pores

    doi: 10.1016/j.bioactmat.2026.02.005

    Figure Lengend Snippet: Channel closure mechanism can be controlled by substrate curvature using scaffolds with well-defined geometries. (a, left) Schematic representation of the in vitro culture setup with collagen scaffold presenting channels of controlled diameter with Ø ≈ 600 μm, Ø ≈ 350 μm and Ø ≈ 150 μm. Monolayer seeding on one side of the biomaterial facilitates migration of cells from one end of the biomaterial. (a, right) SEM image of the microarchitecture (Scale bar 20 μm) and channels within the biomaterial (Scale bars 100 μm). SEM images correspond to the outermost surface of the scaffold. (b) Comparison of template diameter against resulting channel diameter after cross-linking and sterilization of the biomaterial. (c) Representative images of fibroblasts, pre-osteoblasts and endothelial cells within channels of distinct diameters 7 days after seeding. Cell cytoskeleton (F-actin) is depicted in magenta and nuclei in blue. Yellow arrows indicate the direction (arrow angle) and degree of alignment (vector length) for the corresponding region. Scale bar close-up images: 25 μm. (d, left) Degree of channel closure for the investigated channel diameters and cell types. (d, right) Relative degree of tissue alignment for the different channel diameters and cell types. Tissue alignment ranges from 0 (fully isotropic) to 1 (fully anisotropic, dashed line). Tissue across the channel and relative degree of is calculated in the central 50 % of each channel. Data displayed as average with standard deviation. N = 4 scaffolds/cell type. 1 donor/cell type. Scale bars 200 μm (unless otherwise stated).

    Article Snippet: Murine pre-osteoblasts (MC3T3-E1, CRL-2593TM, ATCC) were cultured in alpha modified minimum essential medium with nucleosides (F 0925, Biochrom AG), supplemented with 10 % v/v FBS, 1 % v/v P/S and 1 % v/v GlutaMAX (35050, Gibco®).

    Techniques: In Vitro, Migration, Comparison, Plasmid Preparation, Standard Deviation

    In vitro cell evaluations. (a, b) Fluorescence microscopic images of NIH 3T3 cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.

    Journal: Bioactive Materials

    Article Title: Slippery dopamine–fluoropolymer hybrid surface for improving biliary stent longevity

    doi: 10.1016/j.bioactmat.2026.02.003

    Figure Lengend Snippet: In vitro cell evaluations. (a, b) Fluorescence microscopic images of NIH 3T3 cells stained with a live/dead kit and corresponding quantitative analysis (n = 4) (scale bars, 100 μm). (c) Cytotoxicity analysis with NIT-3T3 cells using CCK-8 kit (n = 4). (d, e) Morphological analysis of NIH 3T3 cells stained for actin (red) and nucleus (blue), with fibroblast aspect ratio analysis (scale bars, 100 μm) (n = 4). (f) Schematic illustration demonstrating the selective application of ELFS coating to the target region. (g, h) Fluorescence images showing selective adhesion of NIH 3T3 and RAW 264.7 cells to ELFS-uncoated region (n = 4) (scale bars, 100 μm). (i, j) Optical images and quantification of adhered colony-forming units (CFUs) on non-coated and ELFS-coated plates after incubation in E. coli and S. aureus suspensions for 24 h (n = 4). (k) Sequential SEM images depicting biofilm formation on non-coated and ELFS- coated stent fragments (n = 3) (scale bars, 0.5 μm). (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001). ns, not significant.

    Article Snippet: The prepared stents were placed on the Transwell insert, and NIH-3T3 fibroblasts (ATCC CRL-1658; 0.5 × 10 5 cells mL −1 ) or human biliary epithelial SNU-1079 cells (Korean Cell Line Bank, KCLB No. 01079; 0.5 × 10 5 cells mL −1 ) were cultured in 2 mL of DMEM supplemented with 10% bovine calf serum and 1% penicillin–streptomycin.

    Techniques: In Vitro, Fluorescence, Staining, CCK-8 Assay, Incubation

    Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a NIH3T3 cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.

    Journal: Journal of Cell Science

    Article Title: OptoLoop – an optogenetic tool to probe the functional role of genome organization

    doi: 10.1242/jcs.264574

    Figure Lengend Snippet: Exploration of optogenetic clustering properties of CRY2. (A) Top panels, time-lapse images of a NIH3T3 cell expressing CRY2high–mCherry activated with a 488-nm microscope laser starting at time t =0 (blue vertical arrow, 1.5 s pulses every 10 s). Scale bars: 5 µm. Bottom panel, coefficient of variation (CV) of fluorescence intensity calculated as the ratio between the nuclear intensity standard deviation and the nuclear intensity mean, presented as relative to the CV at time t =0. Data points corresponding to the images are marked in red. (B) Top panel, protein sequence of the C-terminus of the CRY2 PHR domain and part of the artificial linker used for C-terminal fusions for wild-type CRY2 (CRY2wt) and for CRY2 mutants. The newly generated variant CRY2hiclu is marked in bold. Mutations relative to the CRY2wt sequence are highlighted in gray. Bottom panel, images of NIH3T3 cells expressing CRY2 mutants fused to mCherry, illuminated with 1 s blue light pulses every 10 s for 15 min, and then fixed. The nucleus is delimited with a yellow line. Scale bars: 5 µm. (C) CV calculated from images obtained from NIH3T3 cells expressing CRY2 variants fused to mCherry, illuminated with pulsed blue light for 15 min, and then fixed, plotted as a function of mCherry nuclear intensity. ∼25 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple logistic fits. (D) Time-lapse images of a NIH3T3 cell expressing CRY2hiclu–mCherry activated once with the 488-nm microscope laser for 15 s at time t =0 (marked with a blue arrow). Scale bars: 5 µm. (E) Mean ( n =25) CV calculated from time-lapse images obtained from NIH3T3 cells expressing CRY2olig-mCherry, illuminated with blue light at time t =0, and then kept without blue light. The clustering ( t c ) and declustering times ( t d ) were determined from individual kinetic curves. (F) t c (top panel) and t d (bottom panel) represented as a function of mCherry nuclear intensity. ∼25–40 cells were analyzed per sample (each dot represents one cell). Continuous lines represent simple exponential (clustering) and linear (declustering) fits.

    Article Snippet: NIH3T3 (mouse fibroblasts, ATCC #CRL-1658), U2OS (from human osteosarcoma, ATCC #HTB-96), HeLa (from human cervical adenocarcinoma, ATCC #CRM-CCL-2) and Lenti-X HEK-293T (from human embryonic kidney, cat. #632180 from Takara Bio, Japan) cell lines were cultured in Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) supplemented with 10% (15% for NIH3T3) fetal bovine serum (Gibco, Waltham, MA, USA) plus 100 IU/ml penicillin and 100 μg/ml streptomycin (Gibco, Waltham, MA, USA) at 37°C in a humidified atmosphere with 5% CO 2 .

    Techniques: Expressing, Microscopy, Fluorescence, Standard Deviation, Sequencing, Generated, Variant Assay

    CBNK-EVs rapidly impair ZIKV infectivity and protect host cells. (a) Multiplex immunohistochemical analysis of ZIKV infection in neural progenitor cells. Co-staining for Nestin (neural progenitor cells, green) and ZIKV E protein (red) was performed, and double-positive cells were quantified across six random fields, mean ± SD, ∗P < 0.05. Scale bar, 20 μm. (b) KEGG pathway enrichment bubble plot from RNA-seq of HTR-8/Svneo cells treated with CBNK-EVs (3 × 10 11 particles/mL, 10 μl) and ZIKV (MOI = 1) for 1 h. (c) GSEA analysis of transcriptomic data from HTR-8/Svneo cells. (d) Western blot analysis of ZIKV E and ZIKV NS5 protein levels after co-incubation of ZIKV particles with CBNK-EVs for the indicated time periods (0–30 min) prior to infection. (e) The abundance of NK cell-associated proteins in CBNK-EVs. ∗P < 0.05 (one-way ANOVA).

    Journal: Bioactive Materials

    Article Title: Cord blood natural killer cell-derived extracellular vesicles inhibit Zika virus infectivity through ITGB2/perforin-mediated envelope disruption in vitro and in vivo

    doi: 10.1016/j.bioactmat.2026.01.030

    Figure Lengend Snippet: CBNK-EVs rapidly impair ZIKV infectivity and protect host cells. (a) Multiplex immunohistochemical analysis of ZIKV infection in neural progenitor cells. Co-staining for Nestin (neural progenitor cells, green) and ZIKV E protein (red) was performed, and double-positive cells were quantified across six random fields, mean ± SD, ∗P < 0.05. Scale bar, 20 μm. (b) KEGG pathway enrichment bubble plot from RNA-seq of HTR-8/Svneo cells treated with CBNK-EVs (3 × 10 11 particles/mL, 10 μl) and ZIKV (MOI = 1) for 1 h. (c) GSEA analysis of transcriptomic data from HTR-8/Svneo cells. (d) Western blot analysis of ZIKV E and ZIKV NS5 protein levels after co-incubation of ZIKV particles with CBNK-EVs for the indicated time periods (0–30 min) prior to infection. (e) The abundance of NK cell-associated proteins in CBNK-EVs. ∗P < 0.05 (one-way ANOVA).

    Article Snippet: Vero E6, BHK21, 293T and HTR-8/Svneo cells were purchased from ATCC.

    Techniques: Infection, Multiplex Assay, Immunohistochemical staining, Staining, RNA Sequencing, Western Blot, Incubation

    Perforin mediates the direct virion-disrupting activity of CBNK-EVs. (a) Quantification of perforin and granzyme B in CBNK-EVs by ELISA. (b) Vero-E6 cells were infected with ZIKV (MOI = 1) after pre-incubation of viral particles with 40 ng/L of recombinant perforin or granzyme B for 2 h. ZIKV E protein levels were assessed by Western blot after 24 h, densitometric analysis of the protein bands is shown (right, n = 3). (c) CBNK-EVs were pre-incubated with ZIKV in the presence or absence of 10 mM EGTA, followed by micrococcal nuclease digestion. Protected ZIKV E RNA was quantified by RT-qPCR to assess virion integrity. (d) Western blot analysis of perforin expression in CBNK cells after transfection with the indicated siRNAs. (e) Quantification of perforin levels from (d). (f) Perforin levels in CBNK-EVs collected from control or perforin-knockdown cells, measured by ELISA and normalized to particle count (per 10 11 particles, n = 6). (g) Vero-E6 cells were infected with ZIKV (MOI = 1) that had been pre-incubated with control or perforin-knockdown CBNK-EVs. ZIKV E protein levels were evaluated by Western blot. (h) Analysis of ZIKV E protein levels from (g). (i) Characterization of ITGB2-EVs and control EVs from 293T cells by NTA and TEM. Scale bar, 100 nm. (j) Zeta potential measurements of ITGB2-EVs and control EVs. (k) Western blot analysis of EV markers and ITGB2 expression in ITGB2-EVs and control EVs. (l) Antiviral activity of CBNK-EVs and ITGB2-EVs evaluated by cell-based ZIKV E protein ELISA. Data are presented as mean ± SD (n = 3). ∗P < 0.05, ∗∗P < 0.01, ∗P < 0.001 (one-way ANOVA).

    Journal: Bioactive Materials

    Article Title: Cord blood natural killer cell-derived extracellular vesicles inhibit Zika virus infectivity through ITGB2/perforin-mediated envelope disruption in vitro and in vivo

    doi: 10.1016/j.bioactmat.2026.01.030

    Figure Lengend Snippet: Perforin mediates the direct virion-disrupting activity of CBNK-EVs. (a) Quantification of perforin and granzyme B in CBNK-EVs by ELISA. (b) Vero-E6 cells were infected with ZIKV (MOI = 1) after pre-incubation of viral particles with 40 ng/L of recombinant perforin or granzyme B for 2 h. ZIKV E protein levels were assessed by Western blot after 24 h, densitometric analysis of the protein bands is shown (right, n = 3). (c) CBNK-EVs were pre-incubated with ZIKV in the presence or absence of 10 mM EGTA, followed by micrococcal nuclease digestion. Protected ZIKV E RNA was quantified by RT-qPCR to assess virion integrity. (d) Western blot analysis of perforin expression in CBNK cells after transfection with the indicated siRNAs. (e) Quantification of perforin levels from (d). (f) Perforin levels in CBNK-EVs collected from control or perforin-knockdown cells, measured by ELISA and normalized to particle count (per 10 11 particles, n = 6). (g) Vero-E6 cells were infected with ZIKV (MOI = 1) that had been pre-incubated with control or perforin-knockdown CBNK-EVs. ZIKV E protein levels were evaluated by Western blot. (h) Analysis of ZIKV E protein levels from (g). (i) Characterization of ITGB2-EVs and control EVs from 293T cells by NTA and TEM. Scale bar, 100 nm. (j) Zeta potential measurements of ITGB2-EVs and control EVs. (k) Western blot analysis of EV markers and ITGB2 expression in ITGB2-EVs and control EVs. (l) Antiviral activity of CBNK-EVs and ITGB2-EVs evaluated by cell-based ZIKV E protein ELISA. Data are presented as mean ± SD (n = 3). ∗P < 0.05, ∗∗P < 0.01, ∗P < 0.001 (one-way ANOVA).

    Article Snippet: Vero E6, BHK21, 293T and HTR-8/Svneo cells were purchased from ATCC.

    Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Infection, Incubation, Recombinant, Western Blot, Quantitative RT-PCR, Expressing, Transfection, Control, Knockdown, Zeta Potential Analyzer

    ITGB2 facilitates CBNK-EVs binding to Zika virions and enhances cellular susceptibility to ZIKV. (a) Multicolor immunofluorescence staining of 293T cells transfected with ITGB2, showing co-localization (yellow) between ITGB2 (green) and ZIKV E protein (red). Scale bar: 20 μm. (b) Quantitative analysis of ZIKV E-positive 293T cells from (a). (c) Fluorescence intensity profile along white arrows in (a), indicating sites of ITGB2 and ZIKV E co-localization. (d) 293T cells were transfected with increasing amounts (0.5, 1, 2 μg) of ITGB2 plasmid and infected with ZIKV (MOI = 1) for 1 h. ZIKV RNA levels were measured by qPCR to assess infection susceptibility (mean ± SD, n = 3). (e) Molecular docking model predicting the interaction interface between ITGB2 and ZIKV E protein. (f) Co-IP assay in 293T cells, followed by immunoblotting with anti-ZIKV E protein antibody. (g) CBNK-EVs were pre-incubated with ITGB2 mAb (0, 5, 20 μg/mL) before being applied to Vero-E6 or BHK-21 cells. Cells were then infected with ZIKV (MOI = 1) for 1 h, and antiviral activity was assessed by measuring ZIKV RNA levels at 24 h post-infection. Data are presented as mean ± SD (n = 3 for d and g; n = 6 for b). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (one-way ANOVA).

    Journal: Bioactive Materials

    Article Title: Cord blood natural killer cell-derived extracellular vesicles inhibit Zika virus infectivity through ITGB2/perforin-mediated envelope disruption in vitro and in vivo

    doi: 10.1016/j.bioactmat.2026.01.030

    Figure Lengend Snippet: ITGB2 facilitates CBNK-EVs binding to Zika virions and enhances cellular susceptibility to ZIKV. (a) Multicolor immunofluorescence staining of 293T cells transfected with ITGB2, showing co-localization (yellow) between ITGB2 (green) and ZIKV E protein (red). Scale bar: 20 μm. (b) Quantitative analysis of ZIKV E-positive 293T cells from (a). (c) Fluorescence intensity profile along white arrows in (a), indicating sites of ITGB2 and ZIKV E co-localization. (d) 293T cells were transfected with increasing amounts (0.5, 1, 2 μg) of ITGB2 plasmid and infected with ZIKV (MOI = 1) for 1 h. ZIKV RNA levels were measured by qPCR to assess infection susceptibility (mean ± SD, n = 3). (e) Molecular docking model predicting the interaction interface between ITGB2 and ZIKV E protein. (f) Co-IP assay in 293T cells, followed by immunoblotting with anti-ZIKV E protein antibody. (g) CBNK-EVs were pre-incubated with ITGB2 mAb (0, 5, 20 μg/mL) before being applied to Vero-E6 or BHK-21 cells. Cells were then infected with ZIKV (MOI = 1) for 1 h, and antiviral activity was assessed by measuring ZIKV RNA levels at 24 h post-infection. Data are presented as mean ± SD (n = 3 for d and g; n = 6 for b). ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (one-way ANOVA).

    Article Snippet: Vero E6, BHK21, 293T and HTR-8/Svneo cells were purchased from ATCC.

    Techniques: Binding Assay, Multicolor Immunofluorescence Staining, Transfection, Fluorescence, Plasmid Preparation, Infection, Co-Immunoprecipitation Assay, Western Blot, Incubation, Activity Assay

    Micronucleus test of Kratom leaf extract after 4 h exposure with S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

    Journal: Toxicology Reports

    Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

    doi: 10.1016/j.toxrep.2026.102206

    Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure with S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

    Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

    Techniques:

    Micronucleus test of Kratom leaf extract after 4 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

    Journal: Toxicology Reports

    Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

    doi: 10.1016/j.toxrep.2026.102206

    Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 4 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

    Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

    Techniques:

    Micronucleus test of Kratom leaf extract after 24 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

    Journal: Toxicology Reports

    Article Title: Genotoxicity risk assessment of a 7-hydroxymitragynine-enriched Kratom preparation: An integrated in silico and in vitro approach

    doi: 10.1016/j.toxrep.2026.102206

    Figure Lengend Snippet: Micronucleus test of Kratom leaf extract after 24 h exposure without S9 in TK6 cells. Results are the mean ± SD of 3 independent experiments. Statistical testing with one-way ANOVA and Tukey’s post-hoc test (* p < 0.05).

    Article Snippet: The human B lymphoblastoid cell line (TK6) (CRL-8015; batch No. 70045146), purchased from ATCC, was cultured in RPMI 1640 medium supplemented with 10 % fetal bovine serum (FBS) and 1 % penicillin/streptomycin.

    Techniques: