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. 2016 Feb 17;310(9):F895–F908. doi: 10.1152/ajprenal.00431.2015

Fig. 4.

Fig. 4.

Puromycin aminonucleoside (PAN)-induced glomerulonephritis in Dgke KO mice promotes glomerular endothelial lesions in the absence of significant complement activation. A: light microscopy images of Dgke KO mice glomeruli (left) and WT littermate (right) 3 days after PAN injections. The capillary spaces in some glomeruli of both Dgke KO and WT mice after PAN injection are occluded. B: quantification of the average patent capillary area in glomeruli of WT and Dgke KO mice after PAN injection, obtained by digital image analysis. Values are means ± SD; n = 3 mice per group, with 10 optical fields per mouse. P values were calculated by Student's t-test. C: TEM images of a glomerulus of a Dgke KO mouse (left) and a WT littermate (right) 3 days after PAN injection. Some endothelial cells are swollen and have enlarged nuclei (white asterisk), and some capillaries present loss of endothelial fenestrations (black arrow). Podocytes' foot processes are often conserved in Dgke KO mice treated with PAN (left; arrowheads) and more extensively effaced in WT mice treated with PAN (right; arrowheads). Scale bars = 5 μm. D: average length of podocyte effacement calculated by digital image analysis of TEM images. Values are means ± SD; n = 10 images per mouse, with 4 mice per experimental group. P values were calculated by Student's t-test. E: 24-h urinary ACR of Dgke−/− and WT mice treated with PAN or Veh as a control. Dgke KO mice are partially protected from developing acute proteinuria and recover faster after PAN treatment, compared with WT controls. Values are means ± SD; n = 4 mice per experimental group. P values were calculated by Student's t-test. F: quantification of the number of podocytes per glomerulus in the 4 experimental groups, obtained by morphological analysis of TEM images. Values are means ± SD; n = 5 glomeruli per mouse, with 3 mice per group. P values were calculated by Student's t-test. G: immunofluorescence microscopy images of complement C3 deposition in glomeruli of Dgke KO mice and WT controls 10 days after PAN injection. Glomeruli are highlighted by a dashed line. One single glomerulus was positive in a Dgke KO mouse (arrow). n = 50 glomeruli counted per mouse, 3 mice per group. Scale bars = 100 μm.