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The delivery of both a fluorescence marker (calcein) and antiviral drugs into cells has been achieved owing to a large extent of internalization of cationic liposomes (labelled by Lyssamine-Rhodamine PE or fluorescein-PE) as demonstrated by fluorescent microscopy and quantified by flow cytometry.
Coating had no effect on either the rate or the extent of internalization of particles (data not shown).
Studies have to be conducted with longer incubation times to compare the extent of internalization of these peptides in neuronal cells.
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Cellular uptake was first studied on uncoated particles with different morphology in order to find out if shape effects could only result in different extents of internalization.
The extent of viral internalization was quantified using the colocalization and internalization algorithms embedded in the BioimageXD software [24].
In contrast to NECA-mediated A3-YFP receptor internalization, pretreatment with PTx significantly reduced the extent of HEMADO-stimulated internalization.
We found that in cells with decreased RACK1 expression levels there is an increase in the extent of M2 internalization with over 60% of receptors internalized after 30 minutes compared to almost 45% of receptors internalized in cells with normal levels of RACK1 expression (Fig. 5).
Therefore, exosome internalization by recipient cells appears to be a cell type dependent process, and the extent of exosome internalization likely depends upon the phagocytic abilities of the recipient cell.
We found similar receptor expression levels at time 0 and a significant decrease in both the rate and extent of M2 internalization when RACK1 was overexpressed, with only about 40% of the receptors internalized after 60 minutes compared to almost 70% of receptors internalized in cells overexpressing M2 only.
Apart from a low nanomolar receptor affinity, the extent of peptide internalization is another, potentially even more important factor for efficient targeted tracer uptake.
When studying the impact of serum proteins on cellular uptake, it seems clear that the extent of NP internalization depends greatly on the presence of a corona.
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