633 conjugated anti mouse and anti rabbit secondary antibodies

Cells were fixed with 3% paraformaldehyde and permeabilized with 0.1% Triton X‐100. The secondary antibodies were Alexa Fluor 488‐, 546‐ or 633‐conjugated anti‐mouse and anti‐rabbit secondary antibodies (Molecular Probes, Eugene, OR, USA). Actin was labelled with phalloidin‐AF546 (Molecular Probes). Cells were imaged with a Leica confocal scanner SP2. Nuclei were stained with Topro‐3 (Life Technologies, Waltham, MA, USA) or Hoechst 33258 (Sigma‐Aldrich).
Cell lysates after SDS/PAGE and immunoblotting were incubated with primary antibodies. Secondary antibodies were obtained from Sigma‐Aldrich, and blots were developed with SuperSignal West Dura (Pierce, Waltham, MA, USA).

Cells were fixed with 3% paraformaldehyde and permeabilized with 0.3% Triton X-100. The secondary antibodies were Alexa Fluor 488-, 546-, or 633-conjugated anti-mouse and anti-rabbit secondary antibodies (Molecular Probes). Actin was labeled with phalloidin-AF546 or AF-660 (Molecular Probes). For S phase analysis, cells were grown for 3 h with 50 μM Bromodeoxyuridine (BrdU). After fixation, coverslips were treated with HCl (1% and 2%), quenched with Borate, and processed for immunofluorescence. Cells were imaged with a confocal microscope Leica AOBS SP8 or with Leica AF6000 LX. Nuclei were stained with Hoechst 33258.
Cell lysates after SDS-PAGE and immunoblotting on nitrocellulose (Whatman) were incubated with primary antibodies. HPR-conjugated secondary antibodies were obtained from Cell Signaling, and blots were developed with Super Signal West Dura (Thermo Fisher Scientific). Primary and secondary antibodies were removed by using Restore PLUS Western Blot Stripping Buffer (Thermo Fisher Scientific), according to the manufacturer. Unless otherwise indicated, all the immunoblot figures were representative of at least two biological replicates. The primary and secondary antibodies used in this work are listed in the additional file 7, Table S6.