Immunofluorescence

Cells were fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) for 10 min at room temperature (RT) and permeabilized via incubation in 3% paraformaldehyde with 0.1% Triton in PBS for 5 min. After being blocked with 5% bovine serum albumin (BSA) in PBS for 2 h, samples were incubated at 4°C O/N with monoclonal primary antibodies.

The following commercial antibodies were used: mouse anti-paxillin, BD Bioscience (#610051), dilution: 1:200 (van de Water et al., 2001 ) and rabbit anti-paxillin Tyr118, Cell Signaling (#2541S), dilution: 1:200 (Mekhdjian et al., 2017 ). Secondary antibodies Alexa Fluor 647 chicken anti-rabbit (LifeTechnologies #A-21443) or DyLight 405 donkey anti-mouse (Javkson Immunofluorescence) were used at 1:200 for 1 h at RT.