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. 2021 Nov 26;40(24):e108662. doi: 10.15252/embj.2021108662

Figure 2. Secretion of soluble PD‐L1 from astrocytes.

Figure 2

  1. C6 cells expressing PD‐L1‐myc treated with the γ‐secretase inhibitor DAPT for 18 h. PD‐L1 was detected using a myc antibody.
  2. Immunoblot of HEK293 cells expressing PD‐L1‐myc and presenilin 1 (hPS1) or a presenilin 1 dominant negative mutant (hPS1 D246A).
  3. Evaluation of three independent experiments of B (n = 3, mean + SEM, Student's t‐test, *P = 0.012, t = 3.4803, df = 4).
  4. 18 h 1 μM PMA treatment of C6 cells expressing PD‐L1‐myc. Secretion of PD‐L1 in the conditioned medium (CM) was detected using antibody AF1019.
  5. Mouse astrocytes were incubated for 48 h with 1 μM PMA and 10 μg/ml TNF‐α/100 U/μl IFN‐γ. Lysates and the CM were immunoblotted using antibody AF1019.
  6. Scheme of PD‐L1 secretion: PMA‐induced S2 cleavage results in the formation of sPD‐L1 and a C‐terminal fragment (PD‐L1‐CTF), that is cleaved by γ‐secretase.