fluorescein fitc conjugated lotus tetragonolobus lectin  (Vector Laboratories)

Journal: bioRxiv

Article Title: Life in the fast lane: Functional consequences of male-female dynamic differences in the renal autoregulation of flow

doi: 10.1101/2025.09.12.675896

Figure Lengend Snippet: (A) Intravital two-photon (2P) microscopy was used to quantify single-nephron tubular fluid flow dynamics in the kidney cortex. Representative time-lapse images with Lucifer Yellow (LY) or 70kDa dextran-rhodamine show real-time changes in tubular fluid flow dynamics in the kidney of an anaesthetized 3-month-old male C57BL/6J mouse. 2M dextran-rhodamine was used together with LY to label the circulating plasma. (B) A representative recording of 70kDa dextran-rhodamine fluorescence intensity within a luminal ROI of the early PT segment in an anaesthetized 3-month-old male C57BL/6J mouse. Raw signals were overlaid with a reference sine wave of fixed frequency as indicated. (C) Resting-state magnetic resonance imaging (MRI) was used to detect physiological oscillations at the multi-nephron spatial scale. With a resolution of 0.5×0.5×0.5mm 3 in a rat kidney, single-voxel recordings indicate collective fluid dynamics among about five adjacent nephrons in the renal cortex . The plot on the right represents a single-voxel time series obtained from a cortical region of an anaesthetized 3-month-old male Sprague-Dawley rat. Raw signals were overlaid with a reference sine wave. (D) Transit-time ultrasonic flow measurement was used to measure real-time renal blood flow through the renal artery. Data shown was collected from an anaesthetized 6-month-old male C57BL/6J mouse, previously reported in ref. . The original 1000Hz data was down-sampled to 2Hz and bandpass-filtered to visualize slow rhythms below 0.1 Hz, overlaid with a reference sine wave. (E) Glomerular filtration rate (GFR) can be estimated by fitting the transcutaneous clearance profile of a single intravenous bolus of FITC-sinistrin. Data shown was collected from an anaesthetized 3-month-old male C57BL/6J mouse, previously reported in ref. . The raw data points, with a 0.5 Hz sampling rate, are colored in red. Detrended signals for a 200-second interval are shown on the right, overlaid with a reference sine wave.

Article Snippet: The following antibodies were employed in this study recognizing KIM-1 (kidney injury molecule-1, Rat, 1:500, R&D Systems, MAB1817), FITC-conjugated LTL (Lotis Tetragonolobus Lectin, 1:1000, Vector Laboratories, FL-1321).

Techniques: Microscopy, Clinical Proteomics, Fluorescence, Magnetic Resonance Imaging, Filtration, Sampling

Journal: Clinical and Experimental Nephrology

Article Title: Nephrotic syndrome induces the upregulation of cell proliferation-related genes in tubular cells in mice

doi: 10.1007/s10157-024-02608-1

Figure Lengend Snippet: Mki67 and Foxm1 were upregulated in various tubular cells. a Ki-67 immunostaining performed on 4 μm paraffin kidney sections. The number of Ki-67-positive cells was assessed in a minimum of 20 randomly selected high-power fields per animal (n = 3). **P < 0.01 (unpaired t-test). b Volcano plot of genes that have the ontology term of DNA-binding transcription factor activity (GO:0003700). c mRNA expression of Foxm1 and its downstream targets analyzed by the quantitative reverse-transcription polymerase chain reaction (n = 6). ***P < 0.001 (unpaired t-test). d Multiple in situ hybridization (ISH) procedures performed for Foxm1 and Mki67 . The sections were subjected to immunostaining of tubular markers after ISH. MKi67 -positive cells and Foxm1 -positive cells were counted in a minimum of 20 randomly selected high-power fields per animal (n = 3). **P < 0.01 and ****P < 0.0001 (unpaired t-test). Lotus tetragonolobus lectin (LTL), NKCC2, NCC, and AQP2 were utilized as markers for the proximal tubule, thick ascending limb, distal tubule, and collecting duct, respectively

Article Snippet: After the RNAscope protocol, the sections were subjected to heat-mediated antigen retrieval and stained with a rabbit polyclonal antibody specific to NKCC2 (catalog no.: ab191315, Abcam; diluted 1:400 in PBS), rabbit polyclonal antibody specific to NCC (catalog no.: AB3553, Sigma-Aldrich; diluted 1:400 in PBS), rabbit monoclonal antibody specific to AQP2 (catalog no.: ab199975, Abcam; diluted 1:1000 in PBS), and FITC-conjugated Lotus tetragonolobus lectin (Vector Laboratories) to identify the type of tubules.

Techniques: Immunostaining, Binding Assay, Activity Assay, Expressing, Reverse Transcription, Polymerase Chain Reaction, In Situ Hybridization

Journal: Clinical and Experimental Nephrology

Article Title: Nephrotic syndrome induces the upregulation of cell proliferation-related genes in tubular cells in mice

doi: 10.1007/s10157-024-02608-1

Figure Lengend Snippet: Mki67 and Foxm1 were upregulated in the adriamycin-induced nephrotic syndrome model. a Urine albumin, serum albumin, total cholesterol, and blood urea nitrogen (BUN) levels 14 days after adriamycin injection (n = 8). ***P < 0.001 (unpaired t-test). b mRNA expression of Mki67 and Foxm1 analyzed by quantitative reverse-transcription polymerase chain reaction (n = 8). c Multiple in situ hybridization (ISH) procedures performed for Foxm1 and Mki67 . The sections were subjected to immunostaining for tubular markers after ISH. ***P < 0.001 (unpaired t-test). Lotus tetragonolobus lectin (LTL), NKCC2, NCC, and AQP2 were utilized as markers for the proximal tubule, thick ascending limb, distal tubule, and collecting duct, respectively. d The number of Mki67 -positive cells and Foxm1 -positive cells was counted in a minimum of 20 randomly selected high-power fields per animal (n = 3). **P < 0.01 (unpaired t-test)

Article Snippet: After the RNAscope protocol, the sections were subjected to heat-mediated antigen retrieval and stained with a rabbit polyclonal antibody specific to NKCC2 (catalog no.: ab191315, Abcam; diluted 1:400 in PBS), rabbit polyclonal antibody specific to NCC (catalog no.: AB3553, Sigma-Aldrich; diluted 1:400 in PBS), rabbit monoclonal antibody specific to AQP2 (catalog no.: ab199975, Abcam; diluted 1:1000 in PBS), and FITC-conjugated Lotus tetragonolobus lectin (Vector Laboratories) to identify the type of tubules.

Techniques: Injection, Expressing, Reverse Transcription, Polymerase Chain Reaction, In Situ Hybridization, Immunostaining