Additional file 2 of Mical modulates Tau toxicity via cysteine oxidation in vivo

Additional file 2: Fig. S1 a Verification of the Tau-Mical interaction via immunoblot analysis. Immunoprecipitation of htauFLAG−2N4R using anti-FLAG coated beads and subsequent western blot analysis using an anti-Mical antibody (+ bait). Anti-FLAG coated beads have equally been mixed with a lysate from flies overexpressing Mical in the absence of htauFLAG−2N4R to ensure non-specific binding of Mical to the beads (-bait). b Representative Western blots from head lysates of flies expressing Tau panneuronally compared with similar lysates co-expressing UAS-Mical or a UAS-Mical RNAi transgene probed for 14–3-3 epsilon and Syntaxin. The bars represent the mean ± SEM relative levels of 14–3-3 epsilon upon modulation of Mical levels. c Representative Western blots from head lysates of flies expressing Tau panneuronally compared with similar lysates co-expressing UAS-Mical or a UAS-Mical RNAi transgene probed for dTau and Syntaxin. The bars represent the mean ± SEM relative levels of dTau upon modulation of Mical levels. d Immunoprecipitation of htauFLAG−2N4R using anti-FLAG coated beads and subsequent western blot analysis using an anti-dTau antibody (+ bait). Anti-FLAG coated beads have equally been mixed with a lysate of elavC155-GAL4/+;Ras2-GAL4/+ flies to ensure non-specific binding of dTau to the beads (-bait). e Representative Western blots from head lysates of flies expressing Tau panneuronally compared with similar lysates co-expressing UAS-Mical or a UAS-Mical RNAi transgene probed with the indicated antibodies. Quantifications of four independent biological replicates are shown below in which levels of the phosphorylated protein were normalized using the Syntaxin (Syx) loading control. The normalized level of Tau expressed alone for each quantification was fixed to 1. The bars represent the mean ± SEM relative levels of Tau phosphorylated at the given site upon modulation of Mical levels over that of Tau expressed alone.