Figure 5: Uptake of Curc-NL and TAT-Curc-NL by hCMEC/D3 cell monolayers by CSLM. The localization and distribution of Curc-NL (A, C) and TAT-Curc-NL (B, D) fluorescently labelled with BODIPY-Sm (green fluorescence) within hCMEC/D3 cells. NL did not induce changes in actin staining in the cell monolayer. Fluorescent NL were visualized by CSLM: cells were incubated with Far red-Phalloidin to visualize the actin filaments (purple fluorescence), and nuclear staining was performed by DAPI (blue staining). Curc-NL displayed very low intracellular uptake (A, C). Curc-TAT-NL was more efficiently taken-up (B and D). hCMEC/D3 cells were incubated with (A, B) LAMP-1 to mark late-endosomes and early-lysosomes and with (C, D) EAA1 to stain early endosomes (red staining). Neither Curc-NL nor Curc-TAT-NL co-localize with early endosomes and late endosomes/early-lysosomes. Scale bar=10 μm. Curc=curcuminderivative; NL= nanoliposomes.