Journal: iScience

Article Title: Domain-autonomous MKLP1 modules divergently regulate mitotic fidelity and polyploidy in a holocentric system

doi: 10.1016/j.isci.2026.117149

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Article Snippet: 4′,6-diamino-2-phenylindole (DAPI) , Beyotime Biotechnology, China , Cat#C1002.

Techniques: Transfection, Cloning, Imaging, Bicinchoninic Acid Protein Assay, Western Blot, Plasmid Preparation, Recombinant, Software

Journal: iScience

Article Title: Midline thalamic neurons show early hypersynchrony during absence seizures in mouse models

doi: 10.1016/j.isci.2026.117673

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Article Snippet: DAPI Fluoromount-G , SouthernBiotech, Birmingham, Alabama, USA , Cat#0100–20.

Techniques: Recombinant, Mutagenesis, Software, Analysis, Silicon

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: Nuclei were counterstained with DAPI (Beyotime) for 10 min. All staining was performed at room temperature in the dark.

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

Journal: iScience

Article Title: Proteomic and functional profiling of suspended jejunal enteroids reveals pharmacokinetic fidelity to native intestine

doi: 10.1016/j.isci.2026.117654

Figure Lengend Snippet: Characterization of morphology, size, and shape distribution of differentiated 3D enteroids in suspension (A) Representative micrographs of AO enteroids: bright-field microscopy shows live enteroids in suspension (left), scanning electron microscopy (SEM) reveals a dense microvillar surface (middle), and transmission electron microscopy (TEM) shows the organized microvilli-rich brush border, tight junctions (TJ), and glycocalyx (right). High-magnification brightfield and SEM images (bottom row) correspond to the areas outlined by dotted squares in the upper row. (B) Representative micrographs of BO enteroids: live bright-field microscopy illustrates typical basal-out morphology (left), SEM shows the smooth basal surface (middle), and TEM reveals the microvilli-rich brush border on the lumen-facing apical surface, confirming basal-out polarity (right). High-magnification views (bottom row) correspond to the areas outlined in the upper row. (C) Circularity analysis of AO ( n = 118) and BO enteroids ( n = 94) in suspension from brightfield micrographs. (D) Total cross-sectional surface area at 6 days of suspension culture for AO ( n = 122) and BO ( n = 94) enteroids. (E) Concentrations of Matrigel proteins (% of protein over total acquired protein) present in AO and BO enteroids in suspension cultures after washing. Data from different donors ( n = 4, Hu22-2jej, Hu22-3jej, Hu22-5jej, and Hu18-9jej) are shown as means +SD. (F) Representative confocal images following staining of zonula occludens-1 (ZO-1, green), phalloidin (F-actin; magenta) and nuclei (DAPI; blue), highlighting the continuous monolayer barrier in AO and BO enteroids. Scale bars, 20 μm. (G) Quantification of apical-out and basal-out polarity in enteroid suspensions by confocal microscopy ( n = 100 per population). (A–D, F–G). Donor Hu18-9jej was used throughout. See also ; and .

Article Snippet: VECTASHIELD Vibrance Antifade Mounting Medium with DAPI , Vector Laboratories , Cat #: H-1800–10.

Techniques: Morphology, Suspension, Microscopy, Scanning Electron Microscopy, Transmission Assay, Electron Microscopy, Brightfield, Analysis, Confocal, Staining

Journal: iScience

Article Title: Proteomic and functional profiling of suspended jejunal enteroids reveals pharmacokinetic fidelity to native intestine

doi: 10.1016/j.isci.2026.117654

Figure Lengend Snippet: Fatty acid transport in 3D enteroids in suspension (A) Representative confocal images following staining with DAPI (nuclei; blue), SLC27A4 (solute carrier family 27 member 4, green) and phalloidin (F-actin; magenta), highlighting the expression pattern of the fatty acid transport protein in AO enteroids (donor Hu18-9jej). Scale bars, 20 μm. (B and C) AO and BO enteroids were incubated with fluorescent fatty acid analog C1-BODIPY-C12 for 10 min and imaged live using widefield fluorescence microscopy. The dashed square indicates the region shown at higher magnification. (B) C1-BODIPY-C12 accumulated in fluorescent lipid droplets in AO enteroids. Scale bars, 10 μm. (C) BO enteroids showed intraluminal accumulation of C1-BODIPY-C12. Scale bars, 20 μm. (D) AO enteroids (donor Hu18-9jej) were co-incubated with fluorescent fatty acid analog C1-BODIPY-C12 and LipidTOX-DR for 10 minto visualize lipid droplets. AO enteroids were imaged live using widefield fluorescence microscopy. The dashed square indicates the region shown at higher magnification. Scale bars, 20 μm; inset scale bars, 10 μm. (A–D) Enteroids from donor Hu18-9jej were used. See also .

Article Snippet: VECTASHIELD Vibrance Antifade Mounting Medium with DAPI , Vector Laboratories , Cat #: H-1800–10.

Techniques: Suspension, Confocal, Staining, Expressing, Incubation, Widefield, Fluorescence, Microscopy

Journal: iScience

Article Title: Proteomic and functional profiling of suspended jejunal enteroids reveals pharmacokinetic fidelity to native intestine

doi: 10.1016/j.isci.2026.117654

Figure Lengend Snippet: BCRP expression, localization and activity in AO enteroids in suspension (A) Schematic illustrating BCRP efflux under physiological function, inhibition or absence (e.g., knockout). (B) Representative confocal images following staining with DAPI (nuclei; blue), BCRP (green) and phalloidin (F-actin; magenta), highlighting the expression pattern of the BCRP transporter in AO enteroids (donor Hu18-9jej). Scale bars, 20 μm. (C) BCRP efflux in AO enteroids (donor Hu18-9jej) calculated by PhA accumulation; transporter substrate: PhA, transporter inhibitor: KO143. Results plotted as mean ± SD. Data are shown as triplicates in three independent experiments; data points of each independent experiment denoted as an individual shape.

Article Snippet: VECTASHIELD Vibrance Antifade Mounting Medium with DAPI , Vector Laboratories , Cat #: H-1800–10.

Techniques: Expressing, Activity Assay, Suspension, Inhibition, Knock-Out, Confocal, Staining

Journal: iScience

Article Title: Proteomic and functional profiling of suspended jejunal enteroids reveals pharmacokinetic fidelity to native intestine

doi: 10.1016/j.isci.2026.117654

Figure Lengend Snippet: CYP3A4 metabolic activity in AO enteroids in suspension and PBPK simulation (A) Representative confocal images following staining with DAPI (nuclei; blue), CYP3A4 (green), and phalloidin (F-actin; magenta), highlighting the expression pattern of the CYP3A4 metabolic enzyme in AO enteroids (donor Hu18-9jej). Scale bars, 20 μm. (B) CYP3A4 metabolic activity in AO enteroids (donor Hu18-9jej) over a time period of 120 min. The concentration of the enzyme substrate terfenadine was monitored over time, as well as the formation of its metabolite fexofenadine. Data are shown as mean ± SD. (C) Intrinsic clearance of terfenadine in AO enteroids (donor: Hu18-9jej) and primary enterocytes (donors: Hu22-32jej, Hu23-1jej and Hu23-5jej). Data are shown as median ± IQR, ∗∗∗ p < 0.001 (Mann-Whitney U test). (D) Corrected intrinsic clearance of terfenadine considering the differences in CYP3A4 protein expression in AO enteroids (donor Hu18-9jej) and primary enterocytes (donors: Hu22-32jej, Hu23-1jej and Hu23-5jej. Data are shown as median ± IQR. Mann-Whitney U test showed no statistically significant difference between groups. (E) Simulated mean pharmacokinetics profile of a 60 mg dose of terfenadine in 140 virtual males (20–50 years, black line) with its 5th and 95th percentile profile shown as dashed lines. The green line represents the PBPK model mean prediction for AO enteroids (log concentration). The orange dots depict mean observed data from Okerholm et al. See also ; .

Article Snippet: VECTASHIELD Vibrance Antifade Mounting Medium with DAPI , Vector Laboratories , Cat #: H-1800–10.

Techniques: Activity Assay, Suspension, Confocal, Staining, Expressing, Concentration Assay, MANN-WHITNEY, Drug discovery

Journal: iScience

Article Title: Proteomic and functional profiling of suspended jejunal enteroids reveals pharmacokinetic fidelity to native intestine

doi: 10.1016/j.isci.2026.117654

Figure Lengend Snippet:

Article Snippet: VECTASHIELD Vibrance Antifade Mounting Medium with DAPI , Vector Laboratories , Cat #: H-1800–10.

Techniques: Tissue, Recombinant, Membrane, Gentle, Cell Recovery, Staining, Plasmid Preparation, Saline, Viability Assay, Lysis, Mass Spectrometry, Software