HeLa and U2OS cell lines (American Type Culture Collection) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS). p130Cas−/− and p130Cas-reconstituted cells (provided by S. Cabodi) were cultured in DMEM with 10% FBS and 1% nonessential amino acids. All cell lines were transfected with the indicated plasmids using Lipofectamine 2000, according to the manufacturer’s instructions. For all experiments, cells were mechanically detached in MACS buffer [containing 1× phosphate-buffered saline (PBS) (pH 7.4), 2 mM EDTA, and 0.5% bovine serum albumin] and seeded in DMEM for 30 min before fixation.

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