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: 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, Airyscan imaging reveals integrin α3 localization at foot process peripheries in both low BP and high BP mice 60 minutes post-blebbistatin. Synaptopodin marks central actin cables (green), integrin α3 shown in red. Scale bar: 1 μm. b, Relative fluorescence intensity plots along indicated lines in panel c demonstrate enhanced integrin accumulation in high BP mice. Greater peak-to-valley intensity differences in high BP samples indicate increased peripheral concentration under elevated shear stress. c, Expansion microscopy reveals foot process boundaries in both low and high BP mice. Podocalyxin staining (magenta) clearly identifies peripheries, with integrin α3 (red) accumulation partially lost in some low BP samples. Scale bar: 1 μm. d, Integrated RFI plots from straightened foot processes show differential integrin distribution. While podocalyxin maintains two peaks surrounding central synaptopodin in both groups, integrin α3 shows widened distribution in low GFR samples versus significant peripheral accumulation in high GFR group, confirming stress-dependent redistribution. e, Airyscan imaging of human kidney samples reveals conserved integrin localization patterns. In healthy human glomerulus (left), integrin α3 (red) localizes at foot process peripheries around synaptopodin-marked central actin (green). In minimal change disease (right), integrin α3 accumulates precisely between effaced foot processes, with sarcomere-like structures (SLSs) visible as discontinuous synaptopodin signals (arrows). Scale bar: 1 μm. f, Relative fluorescence intensity plots from human samples demonstrate peripheral integrin accumulation away from central synaptopodin signals in both healthy and diseased tissue, confirming conservation of the stress-responsive redistribution mechanism across species.

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: Imaging, Fluorescence, Concentration Assay, Shear, Microscopy, Staining

Journal: bioRxiv

Article Title: mRNA Therapy for Alport Syndrome

doi: 10.64898/2026.01.20.700554

Figure Lengend Snippet: (A) Composite tile scan of healthy ( Col4a5 WT) mouse kidney sections shows minimal LNP transfection in the kidney. Tile scans are composite images using a 20x objective. (B) Podocalyxin stained glomeruli shows rare LNP transfection. Taken with 40x water immersion objective. (C) Tile scan showing increased glomeruli transfection and GFP expression in Alport kidneys (yellow arrows). (D) Podocalyxin staining shows increased LNP transfection in the glomerulus in Alport kidneys. Taken with 40x water immersion objective.

Article Snippet: The blocking buffer was then removed and replaced with a polyclonal goat anti-mouse podocalyxin primary antibody diluted in blocking buffer at a 1:200 dilution (R&D Systems cat# AF1556) and incubated overnight in a humidified chamber, protected from light, at 4°C.

Techniques: Transfection, Staining, Expressing