Figure 4

DAPK2 is not expressed in fibrotic cells. Representative immunofluorescence of kidney tissue from ten fields analyzed in two wild type (A-C) uninjured and (D-F) chronic cisplatin injured mice shows no DAPK2 staining in the expanded αSMA positive interstitium of injured mice, white arrows indicate DAPK2 positive cells, open arrows indicate αSMA positive cells. (G) Schematic of primary fibroblast isolation. (H) Representative immunofluorescent staining of ten fields of cultured primary fibroblasts for the fibroblast marker vimentin in primary cells at passage 0. Representative immunofluorescent staining of ten fields of cultured primary fibroblasts for DAPK2 in (I) high and (J) low density primary cells at passage 0. (K) Immunoblot of DAPK2 expression in isolated real fibroblasts demonstrating loss expression of DAPK2 after expansion and passage, arrow indicates DAPK2 band, higher band shows β - Tubulin loading control. Scale bars = 50 μm.