For automated microscopy, control or siRNA-silenced cells were seeded in 24-well plates (CELLSTAR®, Greiner-Bio, Stonehouse, UK) before fixation 24 or 48 h post-seeding at 50-70% cell confluency. Cells were permeabilised, unspecific antibody binding was blocked and cells were stained as for regular immunofluorescence. After the secondary antibody staining, cells were washed and then stained with a whole cell stain (Whole Cell Stain Blue, Thermo Fisher) diluted 1:500 in PBS from a DMSO stock solution for 30 min at room temperature. Finally, cells were washed twice with PBS and overlaid with 1 ml fresh PBS per well before imaging or storage at 4°C until imaging.
