Figure 6
EphrinB1/Fc stimulates tyrosine phosphorylation of EphB2 and deletion of the kinase domain (VM) but not the PDZ binding domain eliminates EphB2-induced tau dephosphorylation.
HEK293-tau cells were treated with ephrinB1/Fc for 45 min and then the phosphorylation level of EphB2 was measured by immunoprecipitation with anti-EphB2 antibody and Western blotting probed by PY99 and anti-EphB2 antibody (a, b). HEK293-tau cells were transfected with deletion of PDZ (c, d) or VM (e, f) domain of EphB2 or the control Vector for 24 h and then treated with ephrinB1/Fc for 45 min. The phosphorylation levels of tau at Thr231 and Ser396, or GSK-3β at Ser9, or PI3K at Tyr458/Tyr199 were measured by Western blotting. The phosphorylation levels of tau and the kinases were normalized against the total levels. Data were representative of at least three independent experiments from 9 ~ 12 culture wells for each group and presented as the mean ± SD. *P < 0.05, **P <0.01 versus Vector.
