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. 2021 Sep 24;13(19):4785. doi: 10.3390/cancers13194785

Figure 2.

Figure 2

Expression of target antigens and RevCARs and binding capability of RevTMs. (a) Number of tumor-associated antigens CD33 or CD123 presented on the surface of MOLM-13, Luciferase (Luc) expressing MOLM-13-Luc or OCI-AML3 cells is shown for three individual experiments as mean ± SD. (b) Expression of CD33 and CD123 on MOLM-13 and OCI-AML3 cells was confirmed by flow cytometry after staining with APC-conjugated anti-CD33 mAb or PE-conjugated anti-CD123 mAb. Binding of His tagged RevTMs on AML cells was detected via FITC-conjugated anti-His mAb. (c) Number of RevCAR-E5B9-28/3z or RevCAR-E7B6-28/3z on RevCAR modified T-cells is quantified for three independent T-cell donors and represented as mean ± SD. (d) Expression of RevCAR-E5B9-28/3z or RevCAR-E5B9-28/3z on modified T-cells was confirmed by staining with anti-E5B9 mAb or anti-E7B6 mAb and PE-conjugated anti-mouse-IgG mAb. Binding of indicated RevTMs to RevCAR-E5B9-28/3z or RevCAR-E5B9-28/3z presenting T-cells was detected using PE-conjugated anti-His mAb. (b,d) Stained cells (black lines) and respective controls (grey lines) are presented as histograms. Percentage of positively stained cells is indicated.