Fig. 3.

Wnt-3a induces the docking of B-raf to DVL-2 and activates B-raf to transduce signal to ERK1/2. a Detection of DVL-2 and B-raf in cell lysates of primary chondrocytes treated with Wnt-3a or vehicle after immunoprecipitation with anti-B-raf antibodies. Total cell lysates were analyzed by immunoblot with anti-DVL-2 and anti-B-raf antibodies. β-Actin was used as a loading control (n = 3). b Detection of Flag-tagged DVL-2 and B-raf proteins in cell lysates of primary chondrocytes, transfected with expression vector for Flag-tagged wild-type DVL-2 or Flag-DVL-2(∆361–736) and treated with Wnt-3a or vehicle, after immunoprecipitation with anti-B-raf antibodies. Total cell lysates were analyzed by immunoblot with anti-Flag and anti-B-raf antibodies. β-Actin was used as a loading control (n = 3). c Detection of phosphorylated and total B-raf in primary chondrocytes treated with Wnt-3a or vehicle for different time points. β-Actin was used as a loading control (n = 3). d Detection of phosphorylated and total ERK1/2 and B-raf in primary chondrocytes transfected with either empty vector or wild-type B-raf-expressing vector and treated with Wnt-3a or vehicle. β-Actin was used as a loading control (n = 3). e Detection of phosphorylated and total ERK1/2 and B-raf in primary chondrocytes transfected with either empty vector or with the vector expressing the dead mutant of B-raf (D594V) and treated with Wnt-3a or vehicle. β-Actin was used as a loading control (n = 3). One representative blot of three independent experiments is shown