Fig. 2

Cp 43 reduced tau phosphorylation by TAOK2. a In vitro kinase assays were carried out using GST-TAOK2 (1–314) and/or recombinant htau (2N4R) in the presence or absence of Cp 43 as indicated. Samples were immunoblotted with antibodies to detect tau-pS262/S356 (12E8), tau-pS202/T205/S208 (AT8) or total tau. b HEK293T cells were co-transfected with empty vector (EV), kinase-deficient TAOK2 (1–349, K57A) or TAOK2 (1–349) and htau (2N4R) as indicated, and cell lysates immunoblotted with antibodies to detect, TAOK-pS181, tau-pS202/T205/S208 (AT8), total tau, Myc or GAPDH. c HEK293T cells were co-transfected with empty vector (EV) or TAOK2 (1–349) and htau (2N4R), and treated with or without Cp 43 as indicated. After 24 h, cell lysates were immunoblotted with antibodies to detect tau-pS202/T205/S208 (AT8), total tau, Myc or GAPDH. d Quantitative analysis of the relative levels of tau-pS202/T205/S208 versus total tau in transfected HEK293T cells is shown. The data are normalised to total tau and the bars represent the average ratio ± SEM (n = 4); **p < 0.01, one-way ANOVA followed by multiple comparison with the Holm-Sidak method