Molecular determinants of WNT9b responsiveness in nephron progenitor cells
Fig 2
Effect of Fzd expression on WNT9b responsiveness in M15 cells.
(A) Cross sections of E11.5 embryos displaying both nephric fields were assessed by in situ hybridization using riboprobes for Fzd1-10, except Fzd9 which was unsuccessful for technical reasons. Asterisk (*) marks ureteric buds. Arrow (→) marks cells of the cap mesenchyme. (B) M15 cells were transiently transfected with β-catenin-luciferase reporter (TOPFlash), Renilla-luciferase reporter, Wnt9b-expression vector and various Fzd1-10 expression plasmids in the presence of recombinant RSPO1 (200 ng/ml). TOPFlash to Renilla signal was measured after 48 hours. A one-way ANOVA followed by a Dunnett correction for multiple comparisons was performed. (**) p = 0.0002 (C) M15 cells were transiently transfected with β-catenin-luciferase reporter (TOPFlash), Renilla-luciferase reporter, Wnt9b-expression vector and siRNAs targeting Fzd1, Fzd2 or Fzd5 vs a scrambled negative control siRNA in the presence of recombinant RSPO1 (200 ng/ml). TOPFlash to Renilla signal was measured. A one-way ANOVA followed by a Dunnett correction for multiple comparisons was performed. (*) p = 0.005.