Microtubule and lactate dehydrogenase binding

Microtubules were purified and polymerized from C57 mouse brains, incubated with W-Tau, and analyzed by dot blot using anti-CW-Tau antibodies and electron microscopy. To address the effect of Tau isoforms in microtubule cytoskeleton SH-SY5Y cells were transduced with pWPI vectors expressing the T30 and CW-T30 isoforms and then, were fixed and stained with antibodies against β-tubulin (SIGMA, T4026) and area was measured using FIJI software.

Lactate dehydrogenase (LDH) was incubated with W-Tau at 80ºC, and aggregation was visualized using transmission electron microscopy (JEM1400 Flash Transmission Electron Microscope, Jeol).