Primers for PCR.

<p>Primers for PCR.</p

Gpr48 deficiency altered the expression of Wnt signal inhibitors and the activity of noncanical Wnt signal pathway.

<p>(<b>a</b>) Western blot analysis shows that p-akt and p-GSK3βwere increased while axin2 was decreased in gpr48 KO mice. (<b>b</b>) Western blot analysis depicts the levels of p-JNK, p-c-jun and p-c-fos in the kidneys of Gpr48 KO and WT adult mice.</p

TGF-β1/Smad signaling was not altered in the kidneys of Gpr48 KO mice.

<p>(a) TGF-β1 expression in the kidney of mice was evaluated by RT-PCR. (b) The phosphorylated forms of smad2 and smad3 were analyzed by western blot.</p

Abnormal expressions of extracellular matrix proteins in the kidneys of Gpr48 KO mice.

<p>Some important fibrotic markers are highly expressed in the Gpr48 KO mice compared with the WT mice. (a) Total RNA was extracted and analyzed for the expression of collagenI, III, IV, MMP1,2,9, TIMP1,2 CTGF, FN and alpha-SMA in the kidneys by RT-PCR. (b) Western blot analysis depicted protein level of collagenI, IV, MMP2 and alpha-SMA in the kidneys of mice. (c) The expression of collagen I, IV, MMP2 and FN proteins was studied by immunohistochemistry on Gpr48 KO and WT adult mice (Magnification, 400×).</p

The incidence of multiple cysts of the kidneys in <i>Gpr48^+/+ and Gpr48^−/−</i> mice.

<p>The incidence of multiple cysts of the kidneys in <i>Gpr48^+/+ and Gpr48^−/−</i> mice.</p

Dominant multiple cysts occurred in the kidneys of the Gpr48 KO mice.

<p>The representative morphology of hematoxylin-eosin stains on kidney sections from Gpr48 WT and KO littermates at the adult age. Dominant multiple cysts occurred in the kidneys of the Gpr48 KO mice (A,C,E), whereas the kidneys in the WT mice (B,D,F) were normal. (Magnification, 25× for (A,B), 200× for (C,D), and 400× for (E,F)).</p

Gpr48 deficiency altered the abilities of cell adhesion.

<p>(a) The expressions of ICAM-1, CD44, E-cadherin and Bcl-2 were determined by RT-PCR. The transcript levels of ICAM-1 and CD44 were increased while the expressions of E-cadherin and Bcl-2 were decreased in Gpr48 KO mice compared with the WT mice. (b) Western blot analyses demonstrate PCNA expression is not changed in Gpr48 KO and WT adult mice.</p

The content of blood urea nitrogen in <i>Gpr48^+/+and Gpr48^−/−</i> mice.

<p>All data are expressed as Mean ± SD, n = 15.</p><p>*p<0.05, <b><i>Gpr48^+/+</i></b> mice vs <b><i>Gpr48^−/−</i></b> mice.</p

Gpr48 deficiency inhibited the expressions of PKD1 and PKD2.

<p>(a) The expressions of PKD1 and PKD2 were determined by RT-PCR. The mRNA levels of PKD1 and PKD2 were significantly decreased in Gpr48 KO mice compared with the WT mice. (b) Representative micrographs demonstrated the protein expression of PC1 was markedly lower in Gpr48 KO mice than in the WT adult mice. Kidneys from Gpr48 KO and WT adult mice were stained immunohistochemically for PC-1 protein. <b>(Magnification, 400×)</b>.</p

β-catenin/Wnt signaling was activated in the kidneys of <i>Gpr48^−/−</i> mice.

<p>(a) The expression of β-catenin was analyzed by Western blot. (b) Western analyses of β-catenin in cytolosic and nuclear fractions in the kidney cells of Gpr48 KO and WT mice. (c)RT-PCR results showed the expressions of different Wnt genes in the kidneys of Gpr48 KO and WT adult mice. (d) The expressions of different Fzd genes were evaluated by RT-PCR in the kidneys of Gpr48 KO and WT adult mice. (e) Western blot analysis shows the dramatic increases of Wnt2b, Wnt4b and Wnt7 in Gpr48 KO mice. All data are expressed as Mean ± SD, n = 15. ***p<0.001, ** p<0.01,*p<0.05, KO vs WT.</p