Figure 2.

TNF-α and TGF-β1 cooperate to promote PD-L1 expression during EMT (A) The expression of immune-checkpoint inhibitors as Galectin-9, CEACAM1, PD-L1, PD-L2, Vista before and after induction of EMT on A549 cells was measured by FACS and measurements were independently repeated at least three times. (B) By FACS the changes in the expression of PD-L1 and EPCAM were also confirmed with 5 days of treatment and after the removal of cytokines during the next five days. (C) At last, the changes in the expression of PD-L1 with TNF-α or TGF-β1 alone, as well as with the combination of these cytokines were measured by FACS. (D and E) Expression of PD-L1 protein and mRNA were measured following TNFα/TGF-β1 treatment by ELISA or qRT-PCR. These experiments were repeated three times with similar results.