Fig. 3. Tubular epithelial YAP activation associated with macrophage infiltration and renal MCP-1 upregulation.

A Representative image of double immunofluorescent staining with YAP and F4/80 in Sham and IRI-28d kidneys. B YAP-activated tubules and F4/80-positive infiltrates were calculated from 55 randomly captured images at IRI-28d kidneys. Correlation between YAP-activated tubules and F4/80-positive infiltrates was analyzed (n = 55). Pearson correlation R value and P value are indicated. C Gene expression analysis for chemokine members including CCL and CXCL family in Mst1/2 knockout mice. Gene Expression Omnibus (GEO) datasets supply the RNA-sequencing data of GES95463. D Regulatory elements of the genes encoding MCP-1 from various mammals were reported by Kim et al.[20]. The TEAD-binding element (red bar) is located 100–400 bp upstream of the transcription start site of MCP-1. TSS transcription start site. E Representative western blot analysis of MCP-1 protein expression in kidney lysates from mice 14 and 28 days after IRI (n = 6 mice per group). Sham-operated mice was used as a control (n = 3 mice). F ELISA analysis of MCP-1 levels in serum. G, H Quantitative polymerase chain reaction analysis of MCP-1 (Ccl2) and CCR2 (Ccr2) mRNA expression in kidneys. AU arbitrary units. Scale bar = 50 µm in all images of A. All values are means ± SEM. *p < 0.05, **p < 0.01, and ^#p < 0.001 defined as significant. ns not significant.