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. 2018 Mar 1;25(1):679–689. doi: 10.1080/10717544.2018.1440666

Figure 2.

Figure 2.

Cellular uptake of fluorescein-labeled DNA complexed with DAB-Lf in PC-3 and DU145 cells: (a) Confocal microscopy imaging of the cellular uptake of fluorescein-labeled DNA (2.5 μg/well) complexed with DAB-Lf, DAB or left uncomplexed, after incubation for 24 h with PC-3 and DU145 cells (Blue: nuclei stained with DAPI (excitation: 405 nm laser line, bandwidth: 415–491 nm), green: fluorescein-labeled DNA (excitation: 543 nm laser line. bandwidth: 550–620 nm), red: Alexa Fluor® 647 probe (excitation: 633 nm laser line, bandwidth: 650–690 nm) (Bar: 10 µm). (b) Flow cytometry quantification of the cellular uptake of fluorescein-labeled DNA (5 µg/well) either complexed with DAB-Lf, DAB or left uncomplexed, after incubation for 24 h with PC-3 cells (n = 5). *p < .05 compared with DAB-Lf-DNA. (c) As in B, but with DU145 cells. (d) Flow cytometry quantification of the PC-3 cellular uptake of fluorescein- labeled DNA (5 µg/well) complexed with DAB-Lf, following pretreatment with free Lf (80 µM) and various cellular uptake inhibitors: phenylarsine oxide (‘PhAsO’), filipin (‘Fil.’), colchicine (‘Colch.’) and poly-L-lysine (‘PLys’) (n = 5), *p < .05 compared with DAB-Lf-DNA. (e) Confocal microscopy imaging of the PC-3 cellular uptake of fluorescein-labeled DNA (2.5 μg/well) complexed with DAB-Lf, following pretreatment with free Lf (80 µM) and various cellular uptake inhibitors: phenylarsine oxide (‘PhAsO’), filipin (‘Fil.’), colchicine (‘Colch.’) and poly-L-lysine (‘PLys’) (Blue: nuclei stained with DAPI (excitation: 405 nm laser line, bandwidth: 415–491 nm), green: fluorescein-labeled DNA (excitation: 543 nm laser line. bandwidth: 550–620 nm), red: Alexa Fluor® 647 probe (excitation: 633 nm laser line, bandwidth: 650–690 nm) (Bar: 10 µm). (f) As in d, but with DU145 cells (g) As in e, but with DU145 cells.