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 Experimental Orthopaedics

Article Title: Collagen Type V alpha 1 chain and alpha‐actinin‐3 variants predict knee ligament injury risk in professional football players

doi: 10.1002/jeo2.70724

Figure Lengend Snippet: Heatmap of combined genotype associations with ligament injury. (a) Proportion of players with any knee ligament injury (ACL + MCL) based on combined genotypes of COL5A1 rs10628678 and ACTN3 rs1815739. (b) Proportion of players with ACL injury is based only on the same combined genotypes. Each cell shows the number of injured players relative to the total in that genotype category, with percentages in parentheses. A darker red colour indicates a higher proportion of injuries. ACL, anterior cruciate ligament; MCL, medial collateral ligament.

Article Snippet: Genotyping of the COL5A1 rs12722, COL5A1 rs10628678 (insertion/deletion), ACTN3 rs1815739 and ACE rs4341 polymorphisms was performed using TaqMan single‐nucleotide polymorphism (SNP) Genotyping Assays (Applied Biosystems) on a LightCycler® 480 System (Roche Molecular Systems) or QuantStudio® Real‐Time polymerase chain reaction (PCR) System (Thermo Fisher Scientific).

Techniques: