Fitc tritc conjugated phalloidin

For standard immunofluorescence, 30/40.000 cells were seeded in full medium onto glass coverslips and incubated according to the experiment. At end point, cells were fixed in 4% PFA, permeabilized with 0.2% Triton and then incubated for 1 h in BSA 1%. Incubation with primary antibodies was carried out overnight in humidified chamber. The day after, coverslips were incubated for 1 h with fluorescence-conjugated secondary antibodies, and then for 30’ with DAPI (Sigma Aldrich, St.Louise, Missouri, USA) and FITC-/TRITC-conjugated Phalloidin (Sigma Aldrich, St.Louise, Missouri, USA). Primary and secondary antibodies used for immunofluorescence are available in Supplementary table 1. Images were taken using both the Olympus BH-2 CCD microscope and a Nikon A1-R confocal laser scanning microscope. Volume view with 3D rendering was carried out using the NIS Elements Advanced Research software (Nikon, Shinagawa, Japan). Super resolution imaging was carried out with CrestOptics X-Light V3 Spinning Disk equipped with super resolution module CrestOptics DeepSIM (CrestOptics, Rome, Italy).