Figure EV5. Abatacept immunoprecipitates PD‐L1:CD80 interactions.

• A
  Western blot analysis of Abatacept immunoprecipitates from indicated DG‐75 cell lines, with and without the BS₃ crosslinker. Precipitates were immunoblotted for GFP (CD80/ CD86) and mCherry (PD‐L1) as indicated. Belatacept modulates anti‐PD‐L1 antibody binding but fails to confer PD‐1 Ig detection.
• B
  Concatenated plot of a one in four serial dilution of Belatacept (starting at 20 μg/ml) of CD80 only (top left hand panel) or CD80/PD‐L1 co‐expressing DG‐75 (top right hand panel), with combined titration curves (bottom left hand panel) and EC50 values (bottom right hand values) of Belatacept binding of CD80 only or PD‐L1/CD80 co‐expressing DG‐75. Data are representative of three independent experiments showing mean ± SEM. ns, not significant: paired t‐test.
• C
  Concatenated plot of a one in four serial dilution of Belatacept (starting at 20 μg/ml) followed by PD‐L1 detection using antibody (MIH3 or 29E.2A3 clone) or PD‐1 Ig, all at 1ug/ml.
• D
  Graphical representation showing PD‐L1 detection under conditions used in (C). Data are representative of three independent experiments showing mean ± SEM.