Fig. 4
Dynamics of NP internalization. Extracts of live fluorescence microscopy images of a and b MDA-MB-231, and c MCF-7 cells exposed to: a and c ANANAS-Atto488-cetux10 (15 µg/mL, Atto488 = 1.6 × 10−7 M) or b cetux-Atto488 (15.1 μg/mL, Atto4888 1.4 × 10−7 M) for 10 min at 37 °C in a flow chamber, and then subjected to washing in order to remove unbound nanoparticles. Cells were continuously subjected to low shear flow (0.02 dyn/cm2) and imaged for up to 60 min thereafter. The panels display the superimposed images from the Atto488-related signal (green), the phase contrast and nuclei related one (blue, Hoescht). Size bar 20 µm. Note that during the imaging period the majority of the Atto488-associated nanoparticles move from the cell periphery to the perinuclear region, whereas the majority of the fluorescent signal associated to cetux-Atto48 remains at the cell surface. The entire live imaging videos from which these representative images were extracted are available in Supplementary Movie 1 and Supplementary Movie 2 (Supplementary Movie 3 is for the non-targeted Formulations; Supplementary Movie 4 and Supplementary Movie 5 show the trafficking process of targeted formulation at longer times- between 1 h to 6 h)
