Fig. (9).

Non-canonical Wnt activation of PKC induces PKC phosphorylation of p300. 3T3 cells were treated with Wnt3a, Wnt5a, or Wnt3a+Wnt5a in the presence or absence of PKC inhibitor for 24 h. Nuclear lysates were prepared and immunoblotted for total p300 and for phospho-Ser89 p300 (using anti-Ser-89 PO4 p300 antibody). Although the levels of total p300 remained essentially unchanged, the levels of phospho-Ser89 p300 were markedly increased upon treatment with conditioned media containing Wnt5a, and the phosphorylation could be blocked by small molecule PKC inhibitor (either Go6976 or Ro31-8425). VC, vehicle control. p300 antibody (N-15, Santa Cruz Biotechnology) used to assess total p300.