Figure 4

Primary mouse renal peritubular endothelial cell performance in mono- and co-culture with mouse proximal tubular epithelial cell over 6 days. (A) Co-localization of VE-cadherin (green) and α-SMA (red) positive cells cultured in MPRECs mono-cultures without VEGF. (B) Peritubular endothelial cell and proximal tubular epithelial cell co-culture. In the co-culture system, isolated peritubular endothelial cells were placed in the bottom chamber of fibronectin pre-coated Transwell plates and proximal tubular epithelial cells were seeded onto the polyester inserts (pore size 0.4 μM). Endothelial cell medium was used for the co-culture. (C) After 6 days, primary mouse renal peritubular endothelial cell performance in mono- and co-culture in the presence or absence of VEGF was assessed by FSP-1(green, top panels) staining and co-localization of α-SMA (red) and VE-cadherin (green, bottom panels). Orange represents α-SMA and VE-cadherin double positive areas. (D) Statistical analysis of primary mouse renal peritubular endothelial cell performance after cells were cultured for 6 days. FSP-1 (green) and α-SMA (red) expression in MRPECs was quantified and expressed as a percentage of positive stained cells against total cells. Images presented are representative of at least 5 independent replicate experiments. Data are expressed as mean ± SEM with N = 5 for each group. *P < 0.05, **P < 0.01 vs. respective control. Original magnification was x 200. Cells in this figure were counterstained with DAPI to visualize nuclei (blue).