Rabbit anti α tubulin primary antibodies

Cells were grown on glass coverslips and cultured as described above. The cells were then fixed in 4% paraformaldehyde for 20 min, permeabilised with 0.1% Triton X-100 in PBS for 25 min, and blocked in 10% goat serum for 1.5 hr. To obtain the MT structure, rabbit anti-α-tubulin primary antibodies (1:50, Proteintech, USA) were diluted with PBS, and the coverslips were incubated at 4°C overnight. The coverslips were washed in PBS and then incubated with goat anti-rabbit secondary antibodies conjugated to cyanine 3 (Cy3, 1:100) for 1 hr at 37°C. The cells were imaged using confocal microscopy (TCS-NT, Leica, Wetzlar, Germany).

Immunocytochemistry was performed using a standard procedure. In brief, cells were grown on glass coverslips and cultured as described above. The cells were washed 3 times with PBS (37°C), fixed in 4% paraformaldehyde for 20 min, and permeabilised with 0.1% TritonX-100 for 15 min.Cells were incubated for 1 h in 5% normal goat serum, followed by immunofluorescence staining. To obtain the MTs structure, rabbit anti-α-tubulin primary antibodies (1 : 50, Proteintech) were diluted with PBS, and the coverslips were incubated at 4°C overnight. The coverslips were washed in PBS and then incubated with goat anti-rabbit secondary antibodies conjugated to cyanine 3 (Cy3, 1 : 100, Proteintech) for 1 h at 37°C. To obtain the F-actin structure, rhodamine phalloidin (1 : 50, Sigma-Aldrich) was diluted with PBS, and the coverslips were incubated at 4°C overnight. The cells were imaged using confocal microscopy (Leica).