Fig. 7

TTBK1 induces physical association of CRMP2 and tau in vitro. a SH-SY5Y cells were transfected with plasmids expressing human wild type tau (2N4R), Myc-tagged WT CRMP2 with or without human wild type TTBK1 (0.5 μg each). Cells were treated with 10 μM freshly prepared Aβ42. The cell lysates were immunoprecipitated with anti-Myc or pT514-CRMP2 antibody, followed by immunoblotting of the immunoprecipitated samples with AT8 (anti-pS202/pS205 pTau), Tau-1 (non-phosphorylated tau), Tau46 (total tau) and total CRMP2 (C4G). 5% of input lysate was also blotted for tau (Tau46) and β-actin for loading normalization. b Quantification of AT8 or Tau46-immureactive bands after normalization with co-precipitated pCRMP2 levels (top panel) or pT514 CRMP2level (bottom panel) (N = 3 per group). Results were representative of 3 independent experiments. The error bars indicate SEM. * and ** denote p < 0.05, or 0.01 as determined by two-way ANOVA and multiple comparisons (N = 3 per group). ANOVA: IP-Myc for AT8; F (3, 8) = 133.718 (p < 0.0001), IP-Myc for Tau46; F (3, 8) = 90.657 (p < 0.0001), IP-pT514 CRMP2 for AT8; F (3, 8) = 144.67 (p < 0.0001), IP-pT514 CRMP2 for AT8; F (3, 8) = 84.63 (p < 0.0001)