Extended Data Fig. 2: Cdk5 treatment increases tau phosphorylation.
From: Tau phosphorylation impedes functionality of protective tau envelopes
a. Schematics of sample preparation of bacterial expressed mNeongreen-tau (red), and Cdk5/p35-treated bacterial expressed mNeongreen-tau (blue). b. Mass-spectrometry-determined degree of phosphorylation of Bact-tau (red) or Bact-Cdk5-tau (blue) along the amino acid sequence (shown above the plot), presented as mean ± s.d. (n = 3, 3 independent samples). c. Mass-spectrometry-determined degree of phosphorylation of Bact-tau (red) or Bact-Cdk5-tau (blue) for individual phosphorylation sites, presented as mean ± s.d. (n = 3, 3 independent samples). Phosphorylation sites with no peptide coverage are marked with a black cross. d. Total relative intensities measured for all peptides covering the phosphorylation site, corresponding to data in panels b and c. Data is presented as mean ± s.d. (n = 3, 3 independent samples). e. Western blot of Bact-tau (bacterial cell expressed tau, red) and Bact-Cdk5-tau (Cdk5/p35-treated bacterial cell expressed tau, blue) using total tau antibody (tau-5, top panel) and phospho-specific antibody (pS404, bottom panel). Three independent samples per condition were analyzed from one Western blot. f. Total tau levels (tau-5 intensity) of Bact-tau (red) and Bact-Cdk5-tau samples (blue), presented as mean ± s.d. (n = 3, 3 independent samples). g. Quantification of relative phosphorylation levels, normalized to total tau. Mean Bact-tau (red) was set to 1 and compared Bact-Cdk5-tau levels (blue), presented as mean ± s.d. (n = 3, 3 independent samples). h. Percentage of taxol-stabilized microtubules covered with tau envelopes after 3 min incubation with Bact-tau (red, mean ± s.d., n = 12, 16, 11 independent experiments) or Bact-Cdk5-tau (blue, mean ± s.d., n = 12, 12, 11 independent experiments). Source data for b, c, d, e, f, g and h is available with this manuscript.
