Fig. 6. Mutational analysis of phospho-residues in hTau. Mutational analysis of phospho-residues in hTau. (A) hTau2N4R with the different phospho-serines that are known to be targeted by TTBK1, TTBK2 and MARK1. (B) Western blot analysis of mutant UAS-hTau2N4R transgenes driven by the n-Syb-Gal4;Gal80[ts] driver. hTau2N4R was detected using a polyclonal hTau antibody; Syx was used as loading control (two independent experiments, 20 adult heads each). hTauTTBK1S6A displayed faster migration than the other hTau2N4R variants. (C,D) Lifespan analysis of mutant hTau2N4R strains driven by the n-Syb-Gal4;Gal80[ts] driver. Results are depicted as Kaplan–Meier survival curves (D) (Kaplan and Meier, 1958) and bar graph (C). hTau2N4R and hTauTTBK1S6E did not show any toxicity while hTauMARK1S6E toxicity was significant when compared to control flies (n-Syb-Gal4;Gal80[ts] crossed to the attP landing site strain BL#9750; error bars indicate mean±s.d.; ***P≤0.001; Dunnett's multiple comparison; n=3 independent experiments; ≥96 flies). Josefin Fernius et al. Biology Open 2017;6:1013-1023 © 2017. Published by The Company of Biologists Ltd