A. Expression levels of secreted PD-L1 detected in
trophoblast supernatants by ELLA assay. Supernatants were collected from 100%
confluent Swan 71 trophoblast cells at different times and PD-L1 expression was
quantified by ELLA assay. Note the increase of sPD-L1 in the supernatant in a
time dependent manner.
B. Trophoblast Swan 71 cells were treated with or without
IFN-β (12.5 IU/ml) for 8, 16, 24 and 48h, after which PD-L1 mRNA
expression was determined by qPCR. IFN-β induced a 6-fold increase in
mRNA expression within 8h of treatment and remained higher than the control for
an additional 48h. Data is presented as fold changes between the IFN-β
treated group compared to the non-treated control group.
C. Trophoblast Swan 71 cells were treated with or without
IFN-β (12.5 IU/ml) for 24 and 48h, after which PD-L1 protein expression
was determined in total cell lysate by western blot analysis. Trophoblast cells
constitutively express intracellular PDL-1 protein, but it is not affected by
IFN-β treatment.
D. Trophoblast Swan 71 cells were treated with or without
IFN-β (12. 5 IU/ml) for 24 and 48 hr, after which supernatants were
collected and sPD-L1 protein expression was determined by ELLA assay.
IFN-β treatment of trophoblast cells enhances the concentration of
soluble PD-L1 detected in the supernatants in a time dependent manner. n=6
individual experiments.