Skip to main content
. 2022 Dec 2;10(12):e005543. doi: 10.1136/jitc-2022-005543

Figure 3.

Figure 3

The in vitro bioactivity of BiTP. (A–C) CCK-8 assays to detect the capability of BiTP against TGF-β-inhibited proliferation of TF-1 cells. (D) Luciferase reporter experiments to evaluate Smad-mediated transcriptional activity. Smad-Luc-transfected A549 cells were treated with 20 ng/mL TGF-β1 and antibodies for 24 hours. Then, luminescence was detected. (E) The diagram showing NFAT luciferase reporter system. In the system, Jurkat-1-PD-1-NFAT-Luc and CHO-K1-PD-L1-CD3L were used. The activity of NFAT-Luc could be hampered by PD-1-PD-L1 axis. (F) NFAT luciferase reporter experiments to detect PD-1 signaling. Jurkat-1-PD-1-NFAT-Luc and BiTP were incubated with CHO-K1-PD-L1-CD3L for 6 hours. Then, luminescence was detected. (G) Superantigen stimulation assay assessing the activity of the anti-PD-L1 moiety of BiTP. PBMCs were mixed with antibodies and staphylococcal enterotoxin A (SEA). Four days later, IL-2 concentration in the supernatants was detected. PBMCs, peripheral blood mononuclear cell; TGF-β, transforming growth factor-beta.