Fig. 6

Cp 43 reduced tau phosphorylation on T427 by TAOK2 in primary neurons. a IRES-EGFP and TAOK2-EGFP transfected neurons (7 DIV) were fixed and immunostained with antibodies to detect tau-pT427 (red) and Myc (magenta). b 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-pT123, tau-pT427, total tau or GST. c TAOK2-EGFP transfected neurons (7 DIV) were incubated with or without Cp 43 (30 μM) for 24 h as indicated and immunostained with antibodies to detect tau-pT427 (red) or total tau (tau-5, magenta). d Quantitative analysis of tau-pT427 immunoreactivity in TAOK2-EGFP transfected neurons incubated with or without Cp 43. Bars represent the average immunofluorescence intensity ± SEM of tau-pT427 normalised to total tau. > 65 neurons were analysed for each group (n = 4). *p < 0.05, student t-test. e-f TAOK2-IRES-Tomato and TAOK2-shRNA-EGFP or scrambled-shRNA-EGFP were co-transfected into neurons as indicated and after 72 h cells were fixed and immunostained for TAOK2 (magenta, e) or tau-pT427 (magenta, f). Nuclei were counterstained with DAPI (blue). Representative maximal projection of z-stack images and immunoblots are shown. Scale bar = 10 μm