Skip to main content
. 2020 Jun 27;24(15):8876–8882. doi: 10.1111/jcmm.15512

Figure 2.

Figure 2

Effect of YAP modulation in vivo and in 3D cultures (A) Representative IHC for aquaporin‐2 (AQP2—collecting duct), Tamm‐Horsfall (THP—distal tubules) and megalin (MEG—proximal tubules) on sequential slides of ASO‐treated mice kidneys. The staining shows the presence of dilated tubules and small cysts that are positive for each marker. Scale bar 100 µm. B, Representative periodic acid‐Schiff (PAS) staining of renal tissue from mice treated with scrambled ASO and Yap ASO. Scale bar 200 µm. C, H&E staining of formalin‐fixed, paraffin‐embedded cysts grown in Matrigel after forskolin stimulation (top row). Representative IHC of formalin‐fixed, paraffin‐embedded cysts of forskolin‐treated Wt and mutant mIMCD3 cells stained for YAP (middle row) and TAZ (bottom row). Wild‐type epithelial cells and Pkd1 KO cells grown in Matrigel spontaneously develop cystic structures with a visible lumen and can swell after forskolin stimulation. Yap KO cells and Yap/Pkd1 double KO cells grown in Matrigel showed impaired cyst formation, with the majority of the cells developing tumour‐like mass structures, which did not respond to forskolin stimulation. Only the sporadic cysts that developed a normal lumen before forskolin treatment increased in size after stimulation. For mIMCD3 Yap/Pkd1 KO, cysts from multiple fields of view are shown separated by a white line. White arrowheads indicate cytoplasmic localization of the proteins; black arrowheads indicate nuclear localization of the proteins. Scale bar 50 µm. D, Gene expression (fold‐change) of YAP/TAZ targets at the sacrifice in mice treated with Yap ASO and scrambled ASO. Each symbol represents a mouse. Mean with ± SD. n.s. (not significant) refers to all the genes in the graph, t‐test. E, Quantification of Ki‐67‐positive area. Each symbol represents a mouse. Mean with ± SD. n.s. not significant, t‐test. F, Gene expression (fold change) of WNT pathway targets Axin2 and Myc at the sacrifice in mice treated with Yap ASO and scrambled ASO. Each symbol represents a mouse. Mean with ± SD. ** P < .01, n.s. not significant, t‐test. G, Gene expression (fold‐change) of TGF‐β pathway targets, Acta2, Col1a1, Vim, Fn1, Pai1 and Mmp2, at the sacrifice in mice treated with Yap ASO and scrambled ASO. Each symbol represents a mouse. Mean with ± SD. * P < .05, ** P < .01, t‐test. If no significance is indicated, the comparison is not significant