Figure 3

Effect of CX3CR1-Ala55Thr on Akt phosphorylation-mediated signaling. (a) Subcellular localization of CX3CR1 wild-type (WT) or its Ala55Thr mutant. mCherry-fused CX3CR1 WT or mutant-expressing vectors were transfected into HEK293 cells and cultured for 36 h before immunofluorescent analysis. DIC, differential interference contrast. Scale bar, 25 μm. (b) Ala55Thr mutation in CX3CR1 inhibits Akt phosphorylation upon fractalkine (FKN) treatment. HEK293 cells transfected with an mCherry-conjugated CX3CR1 WT or Ala55Thr-expressing vector were cultured for 36 h, and then the cells were stimulated with the CX3CR1 ligand fractalkine for 15 min. Cell lysates were analyzed by western blotting using the indicated antibodies. The data are expressed as means±s.e.m. and statistical significance was tested with analysis of variance followed by Tukey–Kramer’s multiple comparison test. **P<0.01, *P<0.05. n=5.