HFn–ICG interaction
with TfR1high CCs. (a–d)
Representative flow cytometry plots showing cell binding with 20,
50, and 100 μg/mL of HFn–ICG (light green, purple, and
blue curves, respectively, black curves = control cells) after 2 h
incubation at 4 °C. (e–h) Representative confocal microscopy
images of cells incubated with HFn–ICG for 2 h at 4 °C
to evaluate binding and TfR1 colocalization (blue = cell nuclei stained
with DAPI, cyan = cell membrane, green = HFn–ICG, and purple
= αTfR1 antibody staining). (i–l) Cellular uptake of
HFn–ICG NPs evaluated by IVIS analysis, after incubation at
37 °C for different time points. High TfR1 expression lead to
high binding, diffused colocalization with TfR1 (white spots) and
strong HFn–ICG uptake.