Immunofluorescence Staining of Transfected Cells

Cells were grown on coverslips to ∼50% confluence and transfected with the respective plasmid. Forty-eight hours posttransfection, the cells were fixed with methanol and washed with PBS repeatedly. Cells were made permeable using 0.2% Triton X-100 in PBS for 15 min at 4°C and then washed with PBS. Cells were then incubated with the respective primary and secondary antibody at 4°C in a humified chamber sequentially. The coverslips were then washed and stained with DAPI (4′,6-diamidino-2-phenylindole) and mounted on slides using Vectashield mounting medium (ThermoFisher catalog number NC9265087). Images were taken using a Zeiss LSM 700 confocal laser-scanning microscope and analyzed using ZEN lite software (48 (link)).