Confocal Microscopy

Four different experiments were performed to investigate the effect of the incubation period of the PEGYlated PβAE-447/pDNA nanoparticles on cellular uptake.

The different cell lines (HEK-293, BEAS-2B, and A549) were cultured in 96-wells for 24 h for cell adhesion. Nanocomplexes in culture media were added to the attached cells for different time periods. After the predefined incubation time intervals, the media containing the nanoparticles were replaced by new media, and the plates were incubated for 48 h. Afterward, the culture media was removed, and the cells were incubated for 10 min in the Hoechst dye 33,342 solutions. Immediately before confocal imaging, the media from the wells were aspirated, and the cells were rinsed twice with PBS(1X). Confocal microscopy images were recorded with a confocal laser scanning microscopy (GmbH Wetzlar, Germany).