MCF10A cell culture and transfection

MCF10A cells were grown as described previously (Bazellières et al., 2015 ) and tested negative for mycoplasma contamination. Cells were transfected using the Lipofectamine 3000 transfection kit (Invitrogen) following manufacturer’s instructions using either a pool of three siRNAs (Bazellières et al., 2015 ) or 5 ng of the plasmid (ZO-1-168S→A-FLAG; S168A), a kind gift from the laboratory of Johanna Ivaska (University of Turku, Finland; Tuomi et al., 2009 ). Five days after transfection, cells were trypsinized and used for experiments. S168A plasmid has a point mutation in serine 168 that impairs its binding to α5 and is tagged with the FLAG peptide for identification.