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. 2022 Aug 15;132(16):e156250. doi: 10.1172/JCI156250

Figure 4. Podocyte density, scarring, stress, and ultrastructure.

Figure 4

(A–E) Podocyte density measured by p57 staining (dark blue, C–E) and summarized in A. Each circle represents an individual mouse. Density was lower in aged IgG2a-injected mice compared with young mice and was increased in aged aPD1ab-injected mice. Glomerular scarring was measured by glomerular collagen IV staining (brown, C–E) and is summarized in B. It was higher in IgG2a-injected aged mice compared with young mice and was lowered by aPD1ab. (F–H) The podocyte stress marker desmin (brown) was increased in aged IgG2a-injected mice compared with young and was lower in aged aPD1ab mice. (I–L) The filtration barrier ultrastructure was assessed by expansion microscopy of FLARE-labeled glomeruli, which demonstrated that glomerular basement membrane (GBM) thickness (pink) was significantly increased in aged IgG2a-injected mice (J) compared with young mice (I) and reduced in aged aPD1ab mice (K). Representative images are shown in I–K, and GBM thickness is quantified in L. N, nuclei; E, erythrocytes. (M–P) Podocyte ultrastructure was characterized by the podocyte exact morphology measurement procedure (PEMP). Representative images are shown in M–O, and filtration slit density (FSD) is quantified in P. This analysis shows a significant decrease in FSD in aged IgG2a-injected mice compared with young mice (M, N, and P). Elevation of FSD was observed in aged aPD1ab mice but did not reach significance (O and P). Scale bars: 5 μm (M–O), 10 μm (I–K), and 50 μm (C–H). Statistical analysis was performed by 1-way ANOVA.