FIGURE 5

FIGURE 5

KMO inhibition impacts cell shape, size and focal adhesions in cultured murine podocytes. Mouse podocytes were differentiated on glass cover slides under nonpermissive conditions and then treated with the KMO inhibitor UPF648 or ethanol control at varying concentrations for 48 h. Changes in cell size were assessed by measuring the total cell area in μm² from individual cells across multiple fields of view from two independent experiments. Cell attachment was determined by the number of DAPI⁺ nuclei per field of view (FOV) that are still adherent to the cover slides after KMO inhibition. The cells were photographed at 10X and all nuclei were counted. Thirty photographs were taken for each group, in two independent experiments. (A) Representative images of mouse podocytes after KMO inhibition. Shown are the focal adhesions (paxillin in green) and Actin filaments (phalloidin in red). Photos were taken at 40X. (B) Quantification of changes in the cell size of mouse podocytes after KMO inhibition. Bars represent mean cell area ±95% CI. Photos were taken at 40X. (C) Average number of DAPI‐labeled nuclei from mouse podocytes per FOV. Bars represent the mean cell number per field of view ±95% CI. (D) Apoptosis as measured by Caspase 3/7 activity in mouse podocytes after KMO inhibition, data is presented as a ratio between the luminescence from UPF648‐treated cells relative to the control of the respective time point. Bars indicate the geometric mean of this ratio from at least 3 wells per condition ± geometric SD. Scale bar = 50 μm (**p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001; ANOVA and Dunnett's multiple comparison test or Kruskal–Wallis test and Dunn's multiple comparison).