Journal: bioRxiv
Article Title: VEGF-C overexpression in kidney progenitor cells is a model of renal lymphangiectasia
doi: 10.1101/2023.05.03.538868
Figure Lengend Snippet: A , Schematic representation of the Six2VegfC mouse model where Six2 + nephron progenitor cells overexpress the growth factor VEGF-C which interacts with lymphatic endothelial cells (LECs) to promote lymphangiogenesis during kidney development. B , Representative images of Six2Vegf-C mice and littermate controls (Ctrl). Six2Vegf-C mice appear runted and have enlarged appearing kidneys of varying severity. Scale bar: 1 mm. C, Mouse weight in grams (g) and serum blood urea nitrogen (BUN) demonstrate reduced weight and kidney function in Six2Vegf-C mice. All data are presented as ±SE. Statistical comparisons were made using an unpaired t test. ***P = 0.0001 and ****P < 0.0001. D, Representative images of H&E stained kidney sections from Six2Vegf-C mice and littermate controls at postnatal day 1 (P1), P14, and 1 month of age demonstrating progressive peripelvic cystic lesions. Scale bar: 200 μm (P1), 500 μm (P14), 500 μm (1 month). E, Higher magnificent images of PAS stained kidney sections from Six2Vegf-C mice. Scale bar: 500 μm (4x), 25 μm (40x). F, Immunofluorescence of Six2Vegf-C kidney cystic lesions labeled for podoplanin (PDPN), LYVE1, VEGFR3, endomucin (EMCN), PDGFRB, Lotus tetraglobulus lectin (LTL), NA/K-ATPase, calbindin 1 (CALB1), pan-keratin (KER), cadherin-1 (CHD1). Scale bar: 50 μm. n ≥ 3 animals/group for all representative images.
Article Snippet: Standard methods for immunofluorescence processing were carried out following heat induced antigen retrieval by citrate buffer (0.01M; pH 6.0) using the following antibodies: PDPN (DSHB Cat# 8.1.1), LYVE1 (R&D Cat# AF2125), VEGFR3 (R&D Cat# AF743), NRP2 (R&D Cat# AF567 EMCN (Abcam Cat# ab106100), PDGFRB (R&D Cat# AF1042), LTL ( Lotus tetraglobulus lectin ) (Vector Laboratories Cat# FL-1321), NA/K-ATPase (DSHB Cat# a5), CALB1 (CST Cat# 13176S), pan-Keratin (CST Cat# 4545S), and CDH1 (CST Cat# 3195S).
Techniques: Staining, Immunofluorescence, Labeling