Immunofluorescence microscopy

Immunofluorescence microscopy was performed as previously described^51,58. A549 cells were grown on coverslips coated with gelatin (0.1%). Cells were treated with SS28 (5 μM, 24 h) and were fixed with 2% p-formaldehyde (20 min at RT). After 24 h of the treatment nonspecific antibody binding sites were blocked using FBS following which cells were incubated with antitubulin antibody (1:500) at RT. Coverslips were washed in PBS, incubated in biotinylated antimouse secondary antibody (1:500; RT for 1 h) followed by Streptavidin-FITC (1:200). Finally, cells were mounted with DABCO and images were captured using Zeiss Fluorescence microscope and analyzed using Axio vision software. Colchicine treated cells were used as a positive control.