Fig. 5. Interactions between Wnt11 and Wnt5a. (A) Wnt11-HA or Xnr2-HA mRNA (1 ng) was injected either alone or together with Wnt5a-Myc mRNA (1 ng) in the same cells at the two-cell stage. Immunoprecipitation was performed with anti-Myc antibody. Co-immunoprecipitation shows Xnr2-HA present in the lysate (lanes 1-2) but absent in the anti-Myc IP-complex (lanes 5-6); Wnt11-HA was present in the whole lysates (lanes 3-4) and precipitated with Wnt5a-Myc (lanes 7-8). Arrowhead, IgG band; asterisk, Wnt11-HA band. (B) Schematic description of same cells (SC) and different cells (DC) injected with tagged Wnt11 and Wnt5a mRNAs at the four-cell stage. (C) Co-immunoprecipitation shows Wnt11-HA present in the whole lysates (lanes 1-2) and precipitated with anti-Myc from both SC and DC injections (lanes 3-4). Wnt5a-HA is detected in whole lysates (lanes 5-6) and after anti-GFP co-IP from SC and from DC injections (lanes 7-8). (D) Schematic description of the TOPflash reporter assay after Wnt mRNA overexpression in the oocyte. (E) TOPflash Luciferase activity measure in blastulae derived from control (uninj), Wnt5a-injected (Wn5a 5 pg), Wnt11 injected (Wnt11 5 and 10 pg) or co-injected (Wnt5a 2.5 pg+Wnt11 5 pg) oocytes.