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. 2019 Jul 28;9(19):5595–5609. doi: 10.7150/thno.34669

Figure 2.

Figure 2

In vitro selectivity for EGFR overexpressing cells and subcellular distribution of 111In and Dox. (A) Western Blot characterising the expression level of EGFR in MDA-MB-468 and MCF7 cells. (B) Competition binding experiments of both 111In-EGF-LP and 111In-EGF to MDA-MB-468 cells when treated with increasing concentrations of non-labelled EGF (cold EGF). Incubation 2 h at 4 °C in PBS (n = 3, standard deviation shown, curve fit by nonlinear regression using Graphpad Prism). (C) Uptake of 111In-EGF-LP by EGFR-positive MDA-MB-468 or EGFR-negative MCF7 breast cancer cells with or without the co-incubation of cold EGF. Incubation 2 h at 37 °C (n = 3, standard deviation shown), **** = p<0.00005 using ANOVA with Bonferroni analysis). (D) Intracellular distribution of 111In within MDA-MB-468 and MCF7 cells following exposure to 111In-EGF-LP, incubation for 2 h at 37 °C (n = 3, standard deviation shown, *** = p<0.0005 and ** = p<0.005 using ANOVA with Bonferroni analysis). (E) Visualisation of cellular uptake of rhodamine-containing liposomes by MDA-MB-468 and MCF7 cells using confocal microscopy. Blue = DAPI, red = rhodamine. Images were processed using ImageJ software. Original magnification: 60x (scale = 25 µm).