Journal: Physiological Reports

Article Title: TLR4 signaling cascades leading to increases in albumin permeability, nephrin ectodomain shedding, and apoptosis in MPC ‐5 podocytes

doi: 10.14814/phy2.71147

Figure Lengend Snippet: Exposure to CRX527 for 24 h does not affect the total amounts of podocin (a), synaptopodin (b), or nephrin (c) as analyzed by immunoblot in cultured podocytes. However, CRX527 induced a significant decrease in abundance of the podocyte transcription factor WT‐1 (d). Points on the bar graphs indicate a replicate of the experiment.

Article Snippet: Antibodies against synaptopodin (sc 515842), nephrin (sc 376522), WT‐1 (sc 192), and podocin (sc 518088) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Western Blot, Cell Culture

Journal: Antioxidants

Article Title: Dynamic Regulation of Ferroptosis in a Neonatal Rat Model of Postnatal Hypoxia-Induced Acute Kidney Injury

doi: 10.3390/antiox15050582

Figure Lengend Snippet: Renal injury and associated biomarkers in the renal cortex of a postnatal hypoxia rat model. ( A ) Expression of the hypoxia marker HIF-1α in the renal cortex, quantified by average optical density (AOD), with HIF-1α-positive staining shown as brown coloration in IHC sections. Representative images were obtained at 200× magnification; scale bar = 50 μm. ( B ) Periodic acid–Schiff (PAS) staining of the renal cortex for the assessment of tubular injury scores, followed by quantitative analysis. Images were acquired at 200× magnification; scale bar = 50 μm. ( C ) Expression of the renal injury marker NGAL in the renal cortex, quantified by AOD, with NGAL-positive staining shown as brown coloration in IHC sections. Images were obtained at 200× magnification; scale bar = 50 μm. ( D ) Glomerular expression of nephrin in each group was quantified as the proportion of nephrin-positive area within the glomerulus, with nephrin staining shown as brown coloration in IHC sections. Images were obtained at 400× magnification; scale bar = 50 μm. For all panels, n = 6 per group. Data distribution was assessed using the Shapiro–Wilk normality test to determine whether parametric or non-parametric statistical analyses were applied. Parameters were analyzed using one-way ANOVA followed by the Sidak post hoc test, and are presented as mean ± SEM ( C , D ). Parameters were analyzed using the Kruskal–Wallis H test followed by Dunn’s post hoc test correction, and are presented as median (IQR) ( A , B ). * p < 0.05 versus normoxia groups (N7 and N14); # p < 0.05 versus H14. In the statistical figures, circles represent the N7 group, squares represent the H7 group, triangles represent the N14 group, inverted triangles represent the H14 group, and diamonds represent the H7N7 group. H7: neonatal rats exposed to hypoxia for 7 days after birth; H14: neonatal rats exposed to hypoxia for 14 days after birth; H7N7: neonatal rats exposed to hypoxia for 7 days followed by normoxia for 7 days; N7: neonatal rats maintained under normoxia for 7 days; N14: neonatal rats maintained under normoxia for 14 days; HIF-1α: hypoxia-inducible factor-1α; NGAL: neutrophil gelatinase-associated lipocalin.

Article Snippet: Primary antibodies included rabbit anti-mouse Alox-12 (1:200; ABclonal, Woburn, MA, USA; A14703), ferritin heavy chain (1:3000; ABclonal; A1144), ferritin light chain (1:100; ABclonal; A21758), FGF-23 (1:200; Bioss, Woburn, MA, USA; bs-5768R), HIF-1α (1:500; Proteintech, Rosemont, IL, USA; A23150), Klotho (1:200; Proteintech; 28100-1-AP), NGAL (1:1000; Abcam; ab63929) and nephrin (1:200; A23150; ABclonal).

Techniques: Expressing, Marker, Staining

Journal: medRxiv

Article Title: Nanoscale Podocyte Morphometrics Predict Disease Progression in IgA Nephropathy

doi: 10.64898/2026.03.30.26349728

Figure Lengend Snippet: (a) OCT frozen kidney biopsies were sectioned, cleared and immunolabelled followed by confocal imaging and deep-learning segmentation. (b) Maximum intensity projection confocal image of nephrin visualizing the slit diaphragm network and foot processes. (c) Output of the semantic segmentation of both the slit diaphragm and foot processes. Foot processes are marked in red and the slit diaphragm in green. (d) Output of the instance segmentation resulting in segmentation of each individual foot process in the image. Each foot process is presented in an individual color. (e-g) From the 2 segmentation outputs, the morphometric parameters SDL (e), FP circularity (f), FP area and FP perimeter (g) are extracted. (e) Schematic image of how SDL is measured. The length of the slit (in blue) is divided by the surface area (outlined in white) resulting in a length-density measurement of slit coverage. (f) Schematic image of how FP circularity is measured. From the segmentation of the individual foot processes, the FP circularity is calculated using the formula, circularity = 4 π (area/perimeter−2). The result is a value between 0 and 1 where a perfect circle has a circularity of 1 and a very elongated rectangle approaches 0. (g) Foot process size is calculated both by FP area and by FP perimeter. The FP area is the pixel area of each segmented foot process converted to µm 2 and the FP perimeter is the outline of the foot process with it closed at the base.

Article Snippet: OCT-frozen biopsy specimens were fixed in PFA, cleared in 4% SDS w/ boric acid and fluorescently immunolabelled for nephrin (R&D Systems, cat. no. AF4269) to visualize the slit diaphragm and podocyte foot processes (FPs) based on the protocol described by Unnersjö-Jess et al (details available in Supplement Methods).

Techniques: Imaging