Rabbit acetylated α tubulin

Cells were fixed in 4% paraformaldehyde in PBS for 20 min at RT and permeabilized in methanol for 5 min at −20 °C. Cells were then blocked with PBS containing 4% BSA for 30 min and incubated with the following primary antibody diluted in the blocking solution overnight at 4 °C: rabbit acetylated α-tubulin (1:4000; Cell Signaling Technology, Inc., Danvers, MA, USA), as previously described^{22 (link),33} . Cells were then rinsed in PBS several times before incubation with secondary antibodies: goat anti-rabbit conjugated with Alexafluor 633 (1:2000; Life Technologies, CA, USA) for 30 min at RT. After extensive washing, coverslips were mounted with Vectashield mounting medium (Vector Laboratories Burlingame, CA, USA) with DAPI and then observed at a Leica TCS SP5 confocal microscope (Leica).

Cells were fixed in 4% paraformaldehyde in PBS for 20 min at room temperature (RT) and permeabilized in methanol for 5 min at −20° C. Cells were then blocked with PBS containing 4% BSA for 30 min and incubated with the following primary antibodies diluted in the blocking solution overnight at 4° C: rabbit acetylated α-tubulin (Cell Signaling, Danvers, USA) 1:1000. Cells were then rinsed in PBS several times before incubation with secondary antibodies, goat anti-rabbit conjugated with Alexafluor 633 (1:2000), for 30 min at RT. After extensive washing, coverslips were mounted with Vectashield mounting medium with DAPI (Vector Laboratories Burlingame, CA, USA) and then observed at confocal laser microscope (Leica, Wetzlar, Germany).