Dkk1 inhibited up-regulation of genes by Rspo2 and Wnt3a.

<p>(<b>A</b>) Immunoblot analysis of total β-catenin (cell lysate, 30 µg protein/lane) and soluble β-catenin (GST-E-cadherin pull-down) from C57MG cells incubated for 3 h with BSA or Rspo2 (10 ng/ml) plus Wnt3a (100 ng/ml), with or without 20 min Dkk1 (1 µg/ml) pre-treatment. Western blot of Hsp70 was loading control. (<b>B–D</b>) Quantitative PCR analysis of <i>Axin2</i>, <i>Ahr</i> and <i>CXCR6</i> expression by C57MG cells after 24 h incubation with BSA or Rspo2 plus Wnt3a, with or without Dkk1 pre-treatment [experiment performed in parallel with (A), using the same reagent concentrations]. Results are means +/− S.D. of triplicate measurements from one of four representative experiments.