Figure 5

Reciprocal responses against Wnt in ciliated and non-ciliated cells. We compared induction of canonical and non-canonical Wnt target genes in response to Wnt3a in cultured ciliated and non-ciliated HK2 tubular epithelial cells. The graphs display relative mRNA expression of indicated genes in ciliated versus non-ciliated cells with or without Wnt3a in culture media. (A) Ciliated and non-ciliated HK2 cells were methanol fixed and were labeled with antibodies against acetylated α-tubulin (cilia) and non-phosphorylated β-catenin. Higher expression of active β-catenin was observed in Wnt3a-treated non-ciliated cells. Arrows highlight normal cilia on epithelial cell. Scale bars: 10 μm. (B-E) Expression of canonical Wnt signaling target genes (AXIN2, WISP2, TP53, CCND1) was substantially induced in ciliated cells but not in non-ciliated cells. (F-I) Addition of Wnt3a to culture media induced expression of non-canonical target genes in ciliated cells but not in non-ciliated HK2 cells (CAMK2A, PLCB1, DAAM1, PTK7). Data is presented as means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, n.s. no significance, DAPI, 4′,6-diamidino-2-phenylindole. P values were calculated respective to each sample without treatment.