2.5. Immunocytochemistry Staining

Cultured cells were washed with PBS and fixed in 4% paraformaldehyde, blocked for one hour with 0.2% bovine serum albumin (in 0.1% Trition-X100), and incubated with myosin 4 monoclonal antibody (ThermoFisher Scientific, Waltham, MA, USA) at 4 °C overnight. The following day the cells were washed and incubated with anti-mouse 488 fluorophore conjugated secondary antibody (Sigma, SAB4600387, 77671-1ML-F, 1:1000) in blocking solution for one hour at room temperature. Cells were washed three times in PBS and then imaged using the EVOS microscope system. In all cases, nuclei were counterstained with DAPI.