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. 2013 Jun 13;24(9):1399–1412. doi: 10.1681/ASN.2012050512

Figure 3.

Figure 3.

Wnt4 is expressed in medullary myofibroblasts during fibrosis. (A) After U-IRI, mice are euthanized at indicated time points and cortex and medulla are dissected out of injured kidney. Transcript expression analysis with qPCR indicates that Wnt4 expression is significantly higher than sham controls 7, 14, and 28 days after injury. Cortical Wnt4 expression is significantly lower than medullary expression at days 7, 14, and 28. Cortical Wnt4 expression is significantly higher than sham at day 14 only. (B) Analysis of fibrotic markers αSMA, fibronectin, and collagen 1α1 by qPCR show significant increase at days 3, 7, 14, and 28 after U-IRI. (C) Kidney cortex and medulla samples are prepared from UUO at 5 and 10 days after surgery and processed for qPCR. Wnt4 transcript is significantly higher than sham in the medulla and cortex 5 days and 10 days after UUO. Medulla Wnt4 expression is significantly higher than cortex expression on both day 5 and day 10. (D) Transcripts for fibrotic genes αSMA, fibronectin, and collagen 1α1 are significantly higher on days 5 and 10 after UUO compared with sham controls. (E) Seven days after U-IRI, anti-GFP antibody immunofluorescent colabeling of outer medulla from CLK and U-IRI kidney sections along with anti-laminin (basement membrane marker), anti-αSMA, and anti-PDGFRβ (myofibroblast markers) antibodies. Nuclear GFP appears outside of all tubules and colocalizes with cells positive for αSMA and PDGFRβ. (F) GFP and CD31 (endothelial cell marker) do not colocalize. (G) GFP and F480 (macrophage marker) do not colocalize. (H) GFP does not colocalize with neutrophils and (I) GFP does not colocalize with CD3 (pan T cell marker). Asterisks in A-D indicate analysis by one-way ANOVA comparing groups to respective sham controls (post test, Tukey’s multiple comparison test). Hash marks in A and C indicate analysis by two-way ANOVA comparing between sample types within the same time-point (post test, Bonferroni). *#P<0.05; **##P<0.01; ***###P<0.001. n=3–4 per group for U-IRI and 4–5 per group for UUO. col1α1, collagen 1α1; DAPI, 4',6-diamidino-2-phenylindole. Scale bars, 50 µM.