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. 2023 Aug 28;31(10):2914–2928. doi: 10.1016/j.ymthe.2023.08.018

Figure 2.

Figure 2

Selected hFAP adapters mediate adenoviral transduction of hFAP-expressing human fibroblasts

Flow cytometry analysis for (A) hFAP expression and (B) CAR expression of the human fibroblast D551 cell line upon antibody staining. The HeLa cell line served as a positive control in the CAR expression analysis. (C) Transduction of human fibroblasts by hFAP adapter-retargeted Ad5. Recombinant Ad5 encoding iRFP670 was pre-incubated with the selected “Top 4” hFAP adapters or the E3_5 blocking adapter and analyzed at two different MOIs (PFU/cell) for transduction of hFAP+ D551 cells in comparison with the untargeted Ad5. Transduction levels were determined via cellular expression of iRFP670 detected by flow cytometry. Bars represent mean transduction level of two biological replicates ± SD. Statistics: unpaired t test; ∗p < 0.05, ∗∗p < 0.005; p values are indicated for each sample with respect to the untargeted Ad5. Representative data of two independent experiments are shown. (D) Incubation time-dependent transduction of human fibroblasts by hFAP adapter-retargeted and untargeted Ad5. Recombinant Ad5 encoding TdTomato was pre-incubated with the hFAP adapter no. 6 and analyzed for transduction of hFAP+ D551 cells in comparison with the untargeted Ad5 after 4, 24, and 48 h incubation time of the (retargeted) adenoviral vector with the cells at an MOI of 10 (PFU/cell). Transduction was measured via cellular expression of TdTomato detected by flow cytometry. Representative data of two biological replicates of one experiment are shown.