Journal: Cell Reports Medicine

Article Title: Patient-derived kidney organoids recapitulate ADPKD and facilitate the identification of Rho pathway inhibitors as candidate therapeutics

doi: 10.1016/j.xcrm.2026.102720

Figure Lengend Snippet: Functional maturation and segment-specific differentiation in multi-lineage adult renal organoids (MAROs) (A) Schematic of the albumin uptake assay. Organoids were incubated with albumin-Cy3 for 24 h via the basolateral compartment of the transwell insert, followed by fixation and staining for analysis. (B) Schematic illustration of the experimental strategy for assessing P-glycoprotein (P-gp)-mediated transport in kidney organoids. (C) MAROs were incubated with Albumin-Cy3 for 24 h to assess receptor-mediated endocytosis. The top panel shows the control condition (Substrate only) with significant albumin-Cy3 internalization, while the bottom panel shows reduced uptake in the presence of an inhibitor (substrate + inhibitor). Scale bars, 50 μm. (D) Co-localization of albumin-Cy3 with CUBN and LRP2 in MAROs. Organoids were incubated with Albumin-Cy3 for 24 h, fixed, and stained for co-localization with CUBN/LTL and LRP2/LTL to confirm receptor-mediated endocytosis. Scale bars, 50 μm. (E) Live cell imaging of intracellular accumulation of calcein-AM in kidney organoids treated with the P-gp inhibitor PSC833 or vehicle control (0.2% DMSO), demonstrating P-gp transporter activity. (F) Whole-mount immunofluorescence analysis of calcein-AM accumulation in kidney organoids under the same conditions as (E), stained with ZO-1 (red), calcein (green), and DAPI (blue). Scale bars, 50 μm. (G) RT-qPCR analysis of AQP2 expression, a principal cell marker, in organoids cultured in media of indicated compositions. Gene expression was measured on day 10 post-induction. Data are presented as mean ± SEM. (H) RT-qPCR analysis of ATP6V1B1 , a marker of intercalated cells, in organoids cultured in different media. Expression levels were assessed on day 10 post-induction. Data are shown as mean ± SEM. (I) Relative expression levels of collecting duct (CD)-associated genes in organoids cultured in CD differentiation medium, as determined by RT-qPCR on day 10 post-induction. Data are presented as mean ± SEM. (J) Immunofluorescence staining comparing control organoids (left) and CD-induced organoids (right), showing localization of principal cell marker AQP2 and intercalated cell markers FOXI1 and PENDRIN. Scale bars, 50 μm. (K) Whole-mount co-immunofluorescence of CD-induced organoids, showing spatial co-localization of principal cells (AQP2) and intercalated cells (ATP6V1B1). Scale bars, 50 μm. Quantification data are expressed as mean ± SEM (∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001; and n.s., no statistics, Student’s t test).

Article Snippet: To assess receptor-mediated endocytosis in proximal tubule–like cells, organoids were transferred onto Transwell inserts and exposed to Albumin-Cy3 (MCE Cat. HY-NP027 final concentration 10 μg/mL) added to the basolateral compartment for 24 h at 37°C.

Techniques: Functional Assay, Incubation, Staining, Control, Live Cell Imaging, Activity Assay, Immunofluorescence, Quantitative RT-PCR, Expressing, Marker, Cell Culture, Gene Expression

Journal: Journal of neurosurgery

Article Title: Elucidating the kinetics of sodium fluorescein for fluorescence guided surgery of glioma

doi: 10.3171/2018.4.JNS172644

Figure Lengend Snippet: Explanatory diagram of the blood and normal brain kinetic study. Cohorts of study animals were administered (via tail vein) low-dose NaFL, high-dose NaFl, or pegylated-fluorescein (PEG-fluorescein) at 15, 30, 60, or 120 minutes before euthanasia. Blood and tissue specimens were then harvested and used to analyze the presence of different forms of fluorescein in various tissue/blood compartments. fl= fluorescein. Figure is available in color online only.

Article Snippet: Dilutions of NaFl, human serum-albumin–bound NaFl (MedChem Express) and PEG-fluorescein were used as control samples in the gels.

Techniques:

Journal: Journal of neurosurgery

Article Title: Elucidating the kinetics of sodium fluorescein for fluorescence guided surgery of glioma

doi: 10.3171/2018.4.JNS172644

Figure Lengend Snippet: Quantification of unbound and bound NaFl in the blood circulation. Gel electrophoresis images of NaFl in blood serum at different times after administration are presented in A–G; low-dose administration data are shown in A–C and high dose in D–G. In a given gel panel, the columns represent, from left to right, the following: control containing diluted stock unbound NaFl (NaFl alone), control containing stock NaFl prebound to albumin (NaFl+Albumin), and blood samples from each of 3 mice (m1, m2, and m3 [2 samples were run per mouse]). The dashed lines delineate bound NaFl (above the dotted line) from unbound NaFl (below the dashed line), which were separated by weight in the gel. The percentages of the total fluorescent signal originating from the unbound and bound NaFl, computed from the gel images, are shown in H and I. Table of p values showing the difference in the percentage of unbound fluorescein between low- and high-dose cohorts at 15, 30, and 60 minutes postadministration is shown in J. Figure is available in color online only.

Article Snippet: Dilutions of NaFl, human serum-albumin–bound NaFl (MedChem Express) and PEG-fluorescein were used as control samples in the gels.

Techniques: Nucleic Acid Electrophoresis, Control

Journal: Journal of neurosurgery

Article Title: Elucidating the kinetics of sodium fluorescein for fluorescence guided surgery of glioma

doi: 10.3171/2018.4.JNS172644

Figure Lengend Snippet: Unbound NaFl crosses the BBB and extravasates into normal brain tissue. A: Quantitation of unbound and bound NaFl detected in whole brain extracts calculated from gel electrophoresis (a.u. = arbitrary units). B: Table of p values showing the difference between the amount of unbound fluorescein in normal brain tissue between low- and high-dose cohorts at all corresponding time points (15, 30, and 60 minutes) after administration. C: Representative fluorescence photomicrographs of naïve brain tissue sections 15, 30, 60, and 120 minutes post–NaFl administration. Original magnification ×20; scale bar = 20 μm. The red structures indicate lectin-stained vessels and green fluorescence indicates NaFl (n = 3–4 mice/group).

Article Snippet: Dilutions of NaFl, human serum-albumin–bound NaFl (MedChem Express) and PEG-fluorescein were used as control samples in the gels.

Techniques: Quantitation Assay, Nucleic Acid Electrophoresis, Fluorescence, Staining

Journal: Journal of neurosurgery

Article Title: Elucidating the kinetics of sodium fluorescein for fluorescence guided surgery of glioma

doi: 10.3171/2018.4.JNS172644

Figure Lengend Snippet: Superficial distribution of NaFl in the naive rodent brain. A: Schematic of mouse brain outlining ROIs (olfactory bulb, cerebrum, and cerebellum) analyzed for spatial and temporal distribution of NaFl. B: Representative fluorescence images of ex vivo naïve brains at different times after administration of low- or high-dose NaFl. C: Time-dependent NaFl fluorescence intensity in normal brain regions (n = 3–4 mice/time point/dose). D: Table of p values showing the difference in the amount of superficial fluorescein fluorescence between low- and high-dose experimental cohorts is exhibited. Figure is available in color online only.

Article Snippet: Dilutions of NaFl, human serum-albumin–bound NaFl (MedChem Express) and PEG-fluorescein were used as control samples in the gels.

Techniques: Olfactory, Fluorescence, Ex Vivo

Journal: Journal of neurosurgery

Article Title: Elucidating the kinetics of sodium fluorescein for fluorescence guided surgery of glioma

doi: 10.3171/2018.4.JNS172644

Figure Lengend Snippet: Extravasation of NaFl from internal regions within the naïve brain. A: Representative fluorescence images of axial sections from the naïve midbrain in mice injected with a low or high dose of NaFl at different time points after administration. B: Time-dependent fluorescence intensity in subregions (cortex, parenchyma, and ventricles) of axial cross sections from naïve rodent brains (n = 3–4 mice/time point/dose). C: Table displaying p values from the comparison of fluorescein fluorescence in brain subregions (cortex, parenchyma, and ventricular regions) in low-versus high-dose experimental cohorts. Figure is available in color online only.

Article Snippet: Dilutions of NaFl, human serum-albumin–bound NaFl (MedChem Express) and PEG-fluorescein were used as control samples in the gels.

Techniques: Fluorescence, Injection, Comparison

Journal: Journal of neurosurgery

Article Title: Elucidating the kinetics of sodium fluorescein for fluorescence guided surgery of glioma

doi: 10.3171/2018.4.JNS172644

Figure Lengend Snippet: Pegylated-fluorescein administration results in reduced accumulation in normal brain tissue. A: Agarose gels of PEG-fluorescein in the blood. Similar to Fig. 3, the columns represent, from left to right, the following: control containing diluted stock unbound NaFl (NaFl alone), control containing stock NaFl prebound to albumin (NaFl+Albumin), PEG-fluorescein control, and fluorescence signals from blood samples from each of 3 mice (m1, m2, and m3 (2 samples were run per mouse]). B: Temporal behavior of pegylated-fluorescein in the blood, computed from the gels is displayed. C: Representative fluorescence images of whole brain surface distribution of pegylated-fluorescein and the fluorescence intensity over time in superficial brain regions (olfactory bulb, cerebrum, and cerebellum) for pegylated-fluorescein. D: Fluorescence axial sections of ex vivo brain samples postadministration of PEG-fluorescein and the fluorescence intensity of PEG-fluorescein in the internal brain subregions (cortex, parenchyma and ventricular regions) measured from brain slices. The pegylated form shows much lower transport into normal brain tissue. All scales (color scales and y-axis in quantitative graphs) were set to provide direct comparison to low-dose NaFl cohorts in Figs. 4B and ​andCC and ​and5A5A and ​andB.B. Figure is available in color online only.

Article Snippet: Dilutions of NaFl, human serum-albumin–bound NaFl (MedChem Express) and PEG-fluorescein were used as control samples in the gels.

Techniques: Control, Fluorescence, Olfactory, Ex Vivo, Comparison

Journal: Journal of neurosurgery

Article Title: Elucidating the kinetics of sodium fluorescein for fluorescence guided surgery of glioma

doi: 10.3171/2018.4.JNS172644

Figure Lengend Snippet: Pegylated-fluorescein and NaFl in glioma-bearing brains. A: Representative fluorescence images of axial slices of excised brains 60 minutes after administration of NaFl or PEG-fluorescein (true tumors are outlined with blue dashed line). B: Tumor-to–contralateral normal brain tissue contrast for NaFl and PEG-fluorescein. C: Fluorescence signal intensity in tumor and normal regions. *p < 0.05; **p < 0.005 (n = 4 mice per cohort).

Article Snippet: Dilutions of NaFl, human serum-albumin–bound NaFl (MedChem Express) and PEG-fluorescein were used as control samples in the gels.

Techniques: Fluorescence