Endocytosis pathways involved in (R/W)9-ASPNA-TR cell uptake.

<p>PPT/HeLa cells were incubated with Opti-MEM medium containing 1.5 µM (R/W)9-ASPNA-TR and (A) 25 µg/ml Alexa Fluor 488-transferrin, (B) 0.15 µM Alexa Fluor 488-dextran or (C) 10 µg/ml Alexa Fluor 488-cholera toxin subunit B at 37°C for 2 h and then analyzed. (D) Cells were incubated with 1.5 µM (R/W)9-ASPNA-TR conjugate for 4 h and grown for 18 h in conjugate-free medium. Lysotracker Green DND-26 was then added for 2 h and cells were observed using a Zeiss Axio Observer.Z1 microscope. Left panels show differential interference contrast images, middle panels fluorescence images, and right panels show overlays of the fluorescent images (i.e., merged green and red channels). Scale bar: 20 µm. Extents of co-localisation of (R/W)9-ASPNA-TR conjugate with the following were determined using the Icy imaging software (<a href="http://icy.bioimageanalysis.org/" target="_blank">http://icy.bioimageanalysis.org/</a>): Alexa Fluor 488-transferrin, 45.22±6.92%; dextran, 42±0.29%; cholera toxin, 47.91±0.01%; and Lysotracker Green DND-26, 48.85±5.09%.