Figure 3

Double immunofluorescent staining of FSGS kidneys to Ki-67, α-tubulin and DAPI or double immunofluorescent staining to caspase-3 and DAPI. A. FSGS kidneys: cysts with simple squamous epithelium (CS) filled with colloid content, interstitium (i) and collecting tubules (ct) contain Ki-67-positive cells (arrows)(square). B. α-tubulin visualizes short cilia (arrows) in the cysts and extremely long cilia in collecting tubules (arrows)(square). C. Nuclear DAPI stain. D. Merging of A + B + C shows relationships between proliferating cells (green) and cilia (red). Immunostaining to Ki-67, α-tubulin and DAPI; scale bar 10 μm. E. Distribution of Ki-67 positive cells (%) in CNF and FSGS nephrotic syndrome. Cysts of proximal tubules with simple squamous epithelium (CS) or simple cuboidal epithelium (CC), proximal tubules with apparently normal simple cuboidal/columnar epithelium (PTNC). Data are shown as mean ± SD. Significant differences (Kruskal–Wallis) indicated by ***p < 0.0001. F. FSGS kidneys: caspase-3-positive apoptotic cells (arrows) in proximal tubules (PTNC) and interstitium (i). G. Nuclear DAPI stain. H. Merging of F + G shows green caspase-3- positive apoptotic nuclei and blue non-apoptotic nuclei. Immunostaining to caspase-3 and DAPI, scale bar 10 μm.