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. 2020 Sep 6;9(1):1809926. doi: 10.1080/2162402X.2020.1809926

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© 2020 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Generation of the Myc-CaP/Neu Cell Line.

(a) Schematic for the transduction of the Her-2/neu neoantigen into the androgen-responsive Myc-CaP cell line. (b) The extracellular rat Her-2/neu (neu) cDNA fragment containing the immunodominant MHC-I epitope recognized by the FVB/N-derived T cell clone TCRVβ4 (RNEU_420-429 peptide: PDSLRDLSVF)²⁶ was ligated into the vector pWPXL. (c) Sorting strategy to isolate Myc-CaP cells based on their expression of the rat neu antigen. Myc-CaP cells pre- and post-transduction with lentivirus containing RNEU_420-429 peptide (top,) and pre- and post-sorting for neu-expressing cells (Myc-CaP/Neu; bottom). (d) Fluorescent detection of Her-2/neu in formalin-fixed WT Myc-CaP and Myc-CaP/Neu tumor cells grown on poly-D-Lysine-coated coverslips. Expression of the antigen on tumor cells was evaluated with CF640-labeled αHer-2/neu antibody (red); nuclei were counterstained with DAPI (blue). Scale bar = 100 μm.