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. 2021 Oct 14;11:20725. doi: 10.1038/s41598-021-99707-w

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Quantitative analysis of QD620-IRS binding affinity by confocal imaging and flow cytometry. (A) Comparison of the expression levels of EGFR specific E749-A750del mutation in HCC827, H520, H1975 and H358 cell lines by immunofluorescence (Green color is from Alexa Fluor®488 secondary antibody, blue color from DAPI). (B) There is little uptake of QD620 in the four cell lines. (C) QD620-IRS uptake in HCC827 cells expressing EGFR 19 exon deleted mutation is considerably higher than in H1975, H520 and H358 cells. The binding of QD620-IRS in HCC827 cells was inhibited by application of gefitinib (100 μmol/L) (red circle is from QD620-IRS and blue color from DAPI). All scale bars 10 μm. (D) Flow cytometric analysis of endocytic rates in HCC827, H1975, H520 and H358 cells incubated with QD620-IRS or QD620 1 h. In (E), error bars indicate the mean ± S.D. of data from three separate experiments. ***P < 0.001 vs. HCC827 group.