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: Bioactive Materials

Article Title: Reconstructing the ischemic osteogenic microenvironment through hierarchical scaffolds orchestrating Mg 2+ signaling and neuropilin-1–mediated angiogenesis

doi: 10.1016/j.bioactmat.2026.02.031

Figure Lengend Snippet: Evaluation of angiogenic potential induced by Mg 2+ and NRP-1. ( A ) Scratch assay in three different microenvironments. ( B ) Transwell assay in three different microenvironments. ( C ) Quantitative analysis of wound healing area. ( D ) Quantitative analysis of number of migration cells. ( E ) Immunofluorescence staining of VEGFA and FGF2, with DAPI for nuclear staining and F-actin for cytoskeleton labeling. ( F ) Tube formation evaluation in four different microenvironments. ( G ) Quantitative analysis of number of junction. ( H ) Quantitative analysis of Flu intensity. ( I ) Western-blot analysis of VEGFA, FGF2, and Nr4a1. ( J-L ) Quantitative analysis of relative protein expression of VEGFA, FGF2, and Nr4a1. Data are presented as mean values ± s.d. (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 (one-way ANOVA).

Article Snippet: For the Transwell assay, BMSCs were seeded in the upper chambers of Transwell inserts (8.0 μm pore size; Servicebio, China), while different culture conditions were applied in the lower chambers according to the experimental groups.

Techniques: Wound Healing Assay, Transwell Assay, Migration, Immunofluorescence, Staining, Labeling, Western Blot, Expressing

Journal: Cell Reports Medicine

Article Title: Robust transcriptomic hallmarks targeting intratumor heterogeneity in intrahepatic cholangiocarcinoma

doi: 10.1016/j.xcrm.2026.102708

Figure Lengend Snippet: Distinct immune microenvironment across LIHV-defined subgroups (A) Immune cell abundance comparison across subgroups using the indicated immune analysis tools across four independent iCCA cohorts. Circle color and size represent log 2 fold change and p value, respectively. (B) Boxplots of CD66b + neutrophils, CD68 + macrophages, CD3 + T cells, CD20 + B cells, and αSMA + fibroblasts across subgroups (Mann-Whitney U test). (C) Heatmap of immune signatures and checkpoint expression summarized as mean Z scores per subgroup across four cohorts. ∗FDR < 0.05; ∗∗FDR < 0.01; ∗∗∗FDR < 0.001. (D) Heatmap of Spearman correlations between chemokine expression and neutrophil infiltration. IHC, immunohistochemistry. (E) Boxplot comparing CXCL5 expression across subgroups in the Fu-iCCA cohort (Mann-Whitney U test). (F) Dot heatmap of chemokine gene expression across major cell types in Xue’s scRNA-seq dataset. (G) Boxplots of average CXCL5 expression in tumor cells and macrophages across subgroups (Mann-Whitney U test). (H) Western blot validating CXCL5 knockdown and overexpression efficiency in RBE and HuCCT1 cells. (I) Quantification of migrated neutrophils in transwell assays co-cultured with modified RBE and HuCCT1 cells ( n = 4 replicates per group; mean ± SD; Student’s t test). (J) Western blot analysis of CXCL5 overexpression in KTP cells. (K) Tumor growth curves of mice injected with control or CXCL5-overexpressing KTP cells ( n = 6 per group; mean ± SEM; Student’s t test). (L) Proportion and number of infiltrated neutrophils in control and CXCL5-overexpressing KTP tumors ( n = 6 per group; Student’s t test). ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001. See also and and .

Article Snippet: For migration assay, 3 × 10 5 neutrophils were seeded in the upper chamber of 3-μm pore Transwell inserts (Servicebio, WG3415).

Techniques: Comparison, MANN-WHITNEY, Expressing, Immunohistochemistry, Gene Expression, Western Blot, Knockdown, Over Expression, Cell Culture, Modification, Injection, Control