Sp5 or sp8 confocal laser scanning microscope

For confocal microscopy, cells were seeded on glass coverslips in 24-well plates. The next day, cells were fixed with 3.7% paraformaldehyde solution (Electron Microscopy Science) in DPBS for 10 min at RT and permeabilized with 0.5% Triton X-100 (Sigma-Aldrich) in DPBS for 5 min at RT before blocking with 2% FBS (Thermo Fisher Scientific) in DPBS for 30 min at RT. Cells were then incubated with the corresponding primary antibodies diluted in 2% FBS in DPBS for 2 h at RT. After washing three times with DPBS, cells were incubated with Alexa Fluor-conjugated secondary antibodies (Thermo Fisher Scientific) diluted in 2% FBS in DPBS for 1 h at RT on a plate shaker protected from light. Cell nuclei were counterstained with DAPI (4′,6-diamidino-2-phenylindole; Sigma-Aldrich). For confocal microscopy, stained cells were inversely mounted onto glass microscope slides using ProLong gold antifade mountant (Thermo Fisher Scientific). Images were acquired using an SP5 or SP8 confocal laser scanning microscope (Leica Microsystems) or a DM IL LED inverted microscope (Leica Microsystems). The quantification of the nuclear NP signal was performed using ImageJ software.

For confocal microscopy, cells were seeded on glass coverslips in 24-well plates. The next day, cells were fixed with 3.7% paraformaldehyde solution (Electron Microscopy Science) in DPBS for 10 min at RT and permeabilized with 0.5% Triton X-100 (Sigma-Aldrich) in DPBS for 5 min at RT before blocking with 2% FBS (Thermo Fisher Scientific) in DPBS for 30 min at RT. Cells were then incubated with the corresponding primary antibodies diluted in 2% FBS in DPBS for 2 h at RT. After washing three times with DPBS, cells were incubated with Alexa Fluor-conjugated secondary antibodies (Thermo Fisher Scientific) diluted in 2% FBS in DPBS for 1 h at RT on a plate shaker protected from light. Cell nuclei were counterstained with DAPI (4′,6-diamidino-2-phenylindole; Sigma-Aldrich). For confocal microscopy, stained cells were inversely mounted onto glass microscope slides using ProLong gold antifade mountant (Thermo Fisher Scientific). Images were acquired using an SP5 or SP8 confocal laser scanning microscope (Leica Microsystems) or a DM IL LED inverted microscope (Leica Microsystems). The quantification of the nuclear NP signal was performed using ImageJ software.