Fig. 6: FVB/N-Nos1apEx3-/Ex3- mice develop glomerular matrix deposition, tubular dilation, and ultrastructural changes to the glomerular filtration barrier.
A Periodic acid-Schiff-stained sections of kidneys from 6-month-old Nos1apEx3-/Ex3- and Nos1apEx3-/+ mice were generated. 40x images of glomeruli were evaluated through an automated ImageJ pipeline to determine glomerular matrix deposition in a blinded manner. Both male and female Nos1apEx3-/Ex3- mice showed significantly increased glomerular matrix deposition. 6 control and 7 Nos1apEx3-/Ex3- mice per sex group were analyzed (glomeruli count from left to right: 151, 177, 153, 177). Graph shows dot plots with median bars for each genotype. Two-tailed Mann–Whitney test; ***p < 0.0001; Red triangles, Nos1apEx3-/Ex3-; Black dots, Nos1apEx3-/+ littermate controls; Scale bar μm. B Kidneys were processed at in (A). Representative overview images (stitched from 2X fields) and inset images (20X) are shown for Nos1apEx3-/+ controls and Nos1ap Ex3-/Ex3- mice. Tubular dilation is observed in homozygous mice as quantified in the dot plot (right). Red triangle, n = 12 Nos1apEx3-/Ex3-; Black dots, n = 14 Nos1apEx3-/+ littermate controls. Five 20X fields per mouse for quantification; Two-tailed Mann–Whitney test; ***p = 0.0004. Scale bar 100 μm. C Representative transmission electron microscopy images are shown for the heterozygous control Nos1ap+/Ex3- and homozygous Nos1ap Ex3- /Ex3- mice (5 animals per genotype analyzed). Red arrowheads point to tertiary foot processes. Semi-quantification of podocyte foot process density per µm of glomerular basal membrane (GBM) and quantification of GMB thickness in nm (left and right panel, respectively). Dot plot of all data points shown for each genotype with box plot overlayed representing span of 25 to 75th percentiles, center line 50th percentile, and minima to maxima whiskers. Two-tailed Mann–Whitney test; ***p < 0.0001; Red triangles, Nos1apEx3-/Ex3-; Black dots, Nos1apEx3-/+ littermate controls; Scale bar 2 μm.
