Fig. 1.

Canonical and non-canonical ligands promote Stat3 phosphorylation through LRP5/6 and Ror2 co-receptors, respectively. a HEK293T cells were stimulated with control or Wnt-3a-conditioned medium for the indicated times. Cells were lysed and proteins were analyzed by WB with specific antibodies. LRP5/6, Jnk2 and Stat3 phosphorylation was determined with anti-phospho antibodies against LRP6 (Ser1490), Jnk (Thr183/Tyr185, Thr221/Tyr223) or Stat3 (Tyr705). b A quantification of three independent experiments carried out as in (a) and Suppl. Figure 1a is represented. c HEK293T cells depleted of Ror2 with a specific shRNA were stimulated with control, Wnt3a- or Wnt5a-conditioned medium for 30 min. Cells were lysed and Jnk2 and Stat3 phosphorylation was determined as above. d, e Control, Wnt3a- or Wnt5a-conditioned medium was incubated with a Wnt5a ab or an irrelevant IgG (both 8 μg/ml) (d) for 1 h or was alternatively supplemented with DKK1 or GFP as control (e). HEK293T cells were treated for 30 min with the indicated medium; cell extracts were prepared and analyzed by WB with the indicated antibodies. f Cells were incubated for the indicated times with recombinant Wnt3a (50 ng/ml) and cell extracts analyzed by WB. g Signals from the different experiments performed as in panels (c–e) and Suppl. Figures 1b–d were densitometered and represented. The mean ± SD of three different experiments is shown