dapi  (Vector Laboratories)

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