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. 2021 Jul 23;12:669103. doi: 10.3389/fimmu.2021.669103

Figure 1.

Figure 1

Generation and characterization of CAR macrophages. (A) Vector maps of tested CAR designs and schematics showing the structures of CARs used in the study. Figure created with BioRender. (B) Membrane-bound CAR expression. Forty-eight hours after retroviral transduction, the expression of synthetic receptors on THP-1 cells was detected by staining with an anti-MYC antibody, followed by flow cytometry analysis. Untransduced THP-1 cells were used as a negative control. The histograms shown in black correspond to the isotype controls, whereas the red histograms indicate positive fluorescence. (C) FACS-based phagocytosis of 293T cells or 293T-S target cells by UTD or different CAR macrophages. Statistical significance was calculated with one-way ANOVA with multiple comparisons. (D) Killing of 293T or 293T-S cells by UTD or anti-S CAR macrophages at 24 h assessed with a luciferase-based assay. (E) Flow cytometry analyses of CAR macrophages stained with a biotinylated S protein followed by streptavidin-FITC. The histograms shown in black correspond to the use of isotype controls with streptavidin-FITC, whereas the red histograms indicate positive fluorescence. The results shown represent three (B) independent experiments. Data are the shown as the mean ± s.d. of four independent biological replicates (C–E). P values were derived by one-way ANOVA followed by Tukey’s posttest (C–E). **p<0.01, ***p<0.001, ****p<0.0001. NS, Not Significant. The circles represent individual data.