Journal: International Journal of Molecular Sciences

Article Title: Mannan-Binding Lectin Is Associated with Inflammation and Kidney Damage in a Mouse Model of Type 2 Diabetes

doi: 10.3390/ijms25137204

Figure Lengend Snippet: Deposition of C4 within the glomeruli. ( A ) Fluorescence immunohistochemical representation of C4 and nephrin in the glomerulus from OB ( n = 5) and WT mice ( n = 5). Nephrin (red), C4 (green), DAPI (blue), and a merged picture with all three wavelengths. EVOS with an x40 lens is used, and the scale bar indicated in white is 75 μm. ( B ) Immunohistochemical representation of C4 and nNOS. nNOS (red), C4 (green), DAPI (blue), and a merged picture with all three wavelengths. ×40 lens is used, and the scale bar indicated in white is 75 μm.

Article Snippet: Nephrin: goat anti-mouse nephrin antibody (BioTechne, AF3159, Minneapolis, MN, USA) and nNOS: goat anti-mouse nNOS (GeneTex, GTX89962, Irvine, CA, USA), both dilution 1:50 and secondary, were donkey anti-goat-647 (ThermoFisher, A21447) diluted 1:500 and donkey anti-rabbit-488 (ThermoFisher, A21206) diluted 1:500 when in combination with C4 (OB, n = 5) and WT ( n = 5).

Techniques: Fluorescence, Immunohistochemical staining

Journal: bioRxiv

Article Title: Orthogonal Force Balance Between Contractility and Shear Stress Governs Podocyte Dynamics

doi: 10.64898/2026.01.22.701159

Figure Lengend Snippet: a, Model-predicted integrin stability as a function of location from foot process periphery to center under varying shear stress conditions. Low stress (blue) shows minimal peripheral preference; increasing stress (orange to red) drives progressive peripheral accumulation with central depletion. b, Schematic of predicted integrin redistribution under mechanical stress. Integrins accumulate at foot process peripheries (green) as stress increases, with potential shape changes and edge detachment under excessive loading. c, Airyscan super-resolution imaging validates predicted pattern in healthy mouse glomerulus. Integrin α3 (red) accumulates in gaps between synaptopodin-marked foot processes (green), with nephrin marking slit diaphragms (blue). Scale bar: 1 μm. d, Relative fluorescence intensity (RFI) plot along indicated line in panel c shows integrin α3 peaks (red) localized between synaptopodin peaks (green), confirming peripheral accumulation pattern. e Expansion microscopy (4× expansion) enables single foot process resolution. Podocalyxin (membrane marker, magenta) encapsulates central synaptopodin (green), with integrin α3 (red) co-localizing at periphery. Scale bar: 1 μm. f, Quantitative analysis of straightened foot processes. Integrated RFI plot from all pixels surrounding foot processes shows central synaptopodin peak flanked by two peaks in both podocalyxin and integrin α3 channels, definitively confirming peripheral integrin localization matching model predictions.

Article Snippet: The first and second antibodies used included: Guinea-pig anti-mouse synaptopodin (ARP, 03-GP94-N, Waltham, MA, USA); Rabbit anti-mouse integrin-α3 (BiCell, 10003, St. Louis, MO, USA); Goat anti-mouse nephrin (R&D System, AF3159, Minneapolis, MN, USA); Goat anti-mouse podocalyxin (R&D System, AF1556, Minneapolis, MN, USA); Alexa fluor-488 Donkey anti-guinea-pig secondary (Jackson ImmunoResearch, 706-545-148, West Grove, PA, USA); Alexa fluor-594 Donkey anti-rabbit secondary (Jackson ImmunoResearch, 711-585-152, West Grove, PA, USA); Dylight-405 Donkey anti-rabbit secondary(Jackson ImmunoResearch, 711-475-152, West Grove, PA, USA); and Alexa fluor-647 Donkey anti-goat secondary (Jackson ImmunoResearch, 705-605-003, West Grove, PA, USA).

Techniques: Shear, Imaging, Fluorescence, Microscopy, Membrane, Marker

Journal: PLOS One

Article Title: Podocyte specific knockout of the natriuretic peptide clearance receptor is podocyte protective in focal segmental glomerulosclerosis

doi: 10.1371/journal.pone.0319424

Figure Lengend Snippet: ( 7 A & 7 B) Expression of nephrin and podocin mRNAs were significantly decreased in glomerular preparations from Gq mice compared to controls and expression of both podocyte proteins were preserved by podocyte specific KO of NPRC. (7C & 7D) The pattern of nephrin protein expression was similar to the mRNA results, but the differences were not statistically significant.

Article Snippet: A goat polyclonal antibody to nephrin [ ] (catalog number: AF3159, R&D Systems, Minneapolis, MN), 5.

Techniques: Expressing

Journal: PNAS Nexus

Article Title: Beyond 2D: A scalable and highly sensitive method for a comprehensive 3D analysis of kidney biopsy tissue

doi: 10.1093/pnasnexus/pgad433

Figure Lengend Snippet: Evaluation of the suitability of tissue-clearing techniques for observing glomeruli. A) An analysis flow of a rat-dissected kidney with tissue-clearing techniques. B) Visually transparent rat kidney images treated with the respective tissue-clearing techniques. C) Representative images of 3D glomeruli immunostained with nephrin (green), which were treated with each tissue-clearing technique. Bar = 70 and 20 μm, respectively. D) Representative high-magnification images of 3D glomerulus immunostained with nephrin (green), which were treated with each tissue-clearing technique. Bar = 1 μm. E) 3D constructed images of a nontransparent rat kidney immunostained with nephrin (green). The right panels show each Z-depth image. Bar = 100 and 150 μm, respectively. F) 3D constructed images of a transparent rat kidney treated with CUBIC and immunostained with nephrin (green). The right panels show each Z-depth image. Bar = 100 and 150 μm, respectively.

Article Snippet: The primary antibodies used in this study were goat antimice nephrin antibody (AF3159; R & D Systems, Minneapolis, MN, USA), sheep antihuman nephrin antibody (AF4269; R & D Systems), and rabbit anti-claudin-1 antibody (ab15098; Abcam, Cambridge, MA, USA).

Techniques: Construct

Journal: PNAS Nexus

Article Title: Beyond 2D: A scalable and highly sensitive method for a comprehensive 3D analysis of kidney biopsy tissue

doi: 10.1093/pnasnexus/pgad433

Figure Lengend Snippet: A 3D pathological examination detects the lesions of glomerular diseases with higher sensitivity. A) An analysis flow of anti-GBM nephritis rats. B) Representative glomerular PAS staining images in the vehicle group and anti-GBM group. Bar = 125 and 20 μm. C) Representative glomerular IF images of nephrin (green), claudin-1 (red), and DAPI (blue) in the vehicle group and anti-GBM group. The arrowheads indicate glomerular pathological lesions of crescent formation. Bar = 20 μm. D) Detection ratio of pathological lesions in the vehicle group and anti-GBM group, using conventional 2D analysis. E) The left panels show 3D overall images of glomeruli in anti-GBM group immunostained with nephrin (green), claudin-1 (red), and DAPI (blue). Bar = 20 μm. The middle panel shows combined images of 3D glomeruli. Bar = 10 μm. The right panels show each slice of glomeruli. Bar = 20 μm. The arrowhead indicates glomerular pathological lesions of crescent formation. F) A comparison of the detection ratio of crescent formation in the anti-GBM group between 2D and 3D analyses. * P < 0.01. G) Glomerulus volume in the vehicle group and anti-GBM group. * P < 0.01. H) Representative glomerular IF images of nephrin (green) with high magnification in the vehicle group and anti-GBM group. The lower right panel indicates the overall glomerular images immunostained with nephrin (green), claudin-1 (red), and DAPI (blue). Bar = 1 μm.

Article Snippet: The primary antibodies used in this study were goat antimice nephrin antibody (AF3159; R & D Systems, Minneapolis, MN, USA), sheep antihuman nephrin antibody (AF4269; R & D Systems), and rabbit anti-claudin-1 antibody (ab15098; Abcam, Cambridge, MA, USA).

Techniques: Staining, Comparison

Journal: PNAS Nexus

Article Title: Beyond 2D: A scalable and highly sensitive method for a comprehensive 3D analysis of kidney biopsy tissue

doi: 10.1093/pnasnexus/pgad433

Figure Lengend Snippet: Human kidney samples could be three-dimensionally analyzed through tissue-clearing techniques. A) An analysis flow of a human-removed kidney with tissue-clearing techniques. B) The left panels show 3D images of a 1-mm-sliced human kidney sample treated with CUBIC-L&R and immunostained with nephrin (green). Bar = μm. The right panel shows each Z-stack image. Bar = 150 μm. C) The left panels show 3D overall images of human glomeruli immunostained with nephrin (green), claudin-1 (red), and DAPI (blue). Bar = 20 μm. The right panel shows each plane of 3D-immunostained glomerulus. D) Representative high-magnification images of human glomerulus immunostained with nephrin (green). The lower right panel indicates the overall images. Bar = 1 μm.

Article Snippet: The primary antibodies used in this study were goat antimice nephrin antibody (AF3159; R & D Systems, Minneapolis, MN, USA), sheep antihuman nephrin antibody (AF4269; R & D Systems), and rabbit anti-claudin-1 antibody (ab15098; Abcam, Cambridge, MA, USA).

Techniques:

Journal: PNAS Nexus

Article Title: Beyond 2D: A scalable and highly sensitive method for a comprehensive 3D analysis of kidney biopsy tissue

doi: 10.1093/pnasnexus/pgad433

Figure Lengend Snippet: Paraffin-embedded human kidney samples could be three-dimensionally and noninvasively analyzed through tissue-clearing techniques. A) An analysis flow of a human paraffin-embedded kidney biopsy sample with tissue-clearing techniques. B) A 3D-reconstructed image of the whole human paraffin-embedded kidney biopsy sample immunostained with nephrin (green), claudin-1 (red) and DAPI (blue). Bar = 200 μm. C) A 3D-reconstructed glomerulus of a human paraffin-embedded kidney biopsy sample immunostained with nephrin (green), claudin-1 (red). and DAPI (blue). Bar = 20 μm. D) Representative high-magnification images of human glomerulus on a paraffin-embedded biopsy sample immunostained with nephrin (green). Bar = 1 μm.

Article Snippet: The primary antibodies used in this study were goat antimice nephrin antibody (AF3159; R & D Systems, Minneapolis, MN, USA), sheep antihuman nephrin antibody (AF4269; R & D Systems), and rabbit anti-claudin-1 antibody (ab15098; Abcam, Cambridge, MA, USA).

Techniques: