Fig. 5.

Phospho-analysis of flies expressing hTau^2N4R and Tau kinases. (A,C) Western blot, using Licor, of fly head extracts after 5 days expression of the transgenes at 29°C (two independent experiments, 20 adult heads each). Transgene expression was induced by incubating the flies at 29°C for five days using the n-Syb-Gal4;Gal80[ts] driver. (A) Total hTau^2N4R was detected using a rabbit hTau antibody and phospho-S202 and S205 were detected using mouse monoclonal AT8 antibody. Syx was used as a loading control. (B) Quantification of AT8/total hTau from two similar experiments in A, using Licor software Studio Image. We observed an increase in the AT8/total hTau ratio in flies expressing MARK1-hTau^2N4R and TTBK2-hTau^2N4R-MARK1, when compared to hTau^2N4R alone (error bars indicate mean±s.d.; *P≤0.05; Student's two-tailed t-test). (C) Total hTau was detected using a mouse hTau5 antibody and phospho-S422 was detected using rabbit polyclonal pS422 antibody. Syx was used as loading control. (D) Quantifications of pS422/total hTau from two similar experiments in C, using Licor software Studio Image. We observed an increase in the AT8/total hTau ratio in flies expressing TTBK1-hTau^2N4R-MARK1, when compared to hTau^2N4R alone (error bars indicate mean±s.d.; *P≤0.05; Student's two-tailed t-test).