Journal: eBioMedicine

Article Title: Renal podocyte senescence in human obesity detected by urinary extracellular vesicles

doi: 10.1016/j.ebiom.2025.106116

Figure Lengend Snippet: Increased proportion of senescence- and inflammation-associated uEVs in obesity . (A) The proportion of P16 + uEVs was significantly higher in patients with obesity (OB) (n = 21) compared to healthy volunteers (HV, n = 10), suggesting magnified cellular senescence in obesity. (B) The proportions of P16 + uEVs co-expressing the podocyte marker PODXL was higher in OB, but not those expressing URAT1, UROMODULIN, or PROMININ; (C) Representative flow cytometry dot plots depicting P16 + PODXL + uEVs in HV and OB; (D) The percentage of MCP-1 + uEVs was elevated in OB, indicating an inflammatory renal phenotype; (E) The proportion of MCP-1 + PODXL + uEVs was higher in OB, while no significant differences are observed in MCP-1 + uEVs expressing renal tubular cell markers; (F) Representative flow cytometry plots showing MCP-1 + PODXL + uEVs in HV and OB; (G) The proportion of P16 + MCP-1 + uEVs was higher in OB, indicating the coexistence of senescence and inflammation in renal-derived uEVs; (H) P16 + MCP-1 + PODXL + uEVs were significantly increased in OB, while other triple-positive renal tubular cells were not; (I) Representative flow cytometry plots for P16 + MCP-1 + PODXL + uEVs in HV and OB; ∗P < 0.05, ∗∗∗∗P < 0.0001; uEVs, urinary extracellular vesicles; HV, healthy volunteers; OB, obesity; PODXL, podocalyxin; MCP-1, monocyte chemoattractant protein-1; URAT1, urate transporter-1; UROMODULIN, uromodulin; PROMININ, prominin-1.

Article Snippet: Flow cytometry analysis of isolated EVs was conducted following staining with 0.5 mM Tag-it Violet (TIV) cell-labelling solution (BioLegend, San Diego, CA) at 37 °C for 2 h. To confirm EVs identity, samples were stained with tetraspanin markers CD9 (BioLegend, Cat#312105), CD63 (BioLegend, Cat#353007), and CD81 (BioLegend, Cat#349503), in addition to cell-specific antibodies, including p16 (Novus, Cat#NB200-174), MCP-1 (ThermoFisher, Cat#MA5-17040), podocalyxin (PODXL; ThermoFisher, Cat#MA5-15846), nephrin (ThermoFisher, Cat#MA5-41644), urate transporter-1 (URAT1; Creative Diagnostics, Cat#DCABH-7215), uromodulin (Novus, Cat#NBP1-50321), and prominin-1 (R&D, Cat#MAB2024).

Techniques: Expressing, Marker, Flow Cytometry, Derivative Assay

Journal: Renal Failure

Article Title: Establishment and optimization of a novel mouse model of hyperuricemic nephropathy

doi: 10.1080/0886022X.2024.2427181

Figure Lengend Snippet: Primer sequences for qPCR.

Article Snippet: Anti-E-cadherin and anti-urate transporter 1 (URAT1) antibodies were purchased from Proteintech (Chicago, IL, USA).

Techniques:

Journal: Renal Failure

Article Title: Establishment and optimization of a novel mouse model of hyperuricemic nephropathy

doi: 10.1080/0886022X.2024.2427181

Figure Lengend Snippet: The expression of uric acid reabsorption transporters in HN mice. (A) mRNA levels of Urat1 and Glut9 in HN mice ( n = 8). (B, C) URAT1 protein expression in Western blot analyses ( n = 3). Data are presented as the mean ± SEM. Groups were compared using one-way ANOVA. * indicates a significant difference between the NC group and HN groups: * p < 0.05, ** p < 0.01. Glut9, glucose transporter 9; HN, hyperuricemic nephropathy; Hx, hypoxanthine; NC, normal control; PO, potassium oxonate; Urat1, urate transporter 1.

Article Snippet: Anti-E-cadherin and anti-urate transporter 1 (URAT1) antibodies were purchased from Proteintech (Chicago, IL, USA).

Techniques: Expressing, Western Blot, Control