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. 2019 Feb 15;116(10):4326–4335. doi: 10.1073/pnas.1819473116

Fig. 1.

Fig. 1.

ROS regulate PD-L1 expression and secretion of immunosuppressive cytokines in BMDMs. (A) Pdl1 mRNA levels in BMDMs obtained from FVB mice (n = 4 per group). Cells were left untreated (Ctrl) or treated for 24 h with BSO (200 μM) ± NAC (1 mM). (B) Gclc and Gclm mRNA levels in BMDMs treated as in A. n = 4 per group. (C and D) Percentage of PD-L1 positive BMDMs gated on live CD45+CD11b+F4/80+ within CD206+MHC-II− (C) or CD206−MHC-II+ (D) populations. Cells were isolated from n = 3 FVB mice and analyzed after being treated as in A. (E) Levels of indicated chemokines in the supernatant of BMDMs, as determined by cytokine antibody array after treatment with BSO (200 μM) ± NAC (1 mM) for 24 h. Values are the mean of biological duplicates and are represented as ratio to untreated control. (F) Pdl1 mRNA levels in BMDMs that were exposed to DMSO (Ctrl) or paclitaxel (100 nM) ± NAC (1 mM) for 24 h. n = 4 per group. (G) Gclc and Gclm mRNA levels in BMDMs treated as in F. (H and I) Percentage of PD-L1 positive BMDM gated on live CD45+CD11b+F4/80+ within CD206+MHC-II− (H) or CD206−MHC-II+ (I) populations. Cells were isolated from n = 3 mice treated as in F. (J) Levels of indicated chemokines in the supernatant of BMDMs as determined by cytokine antibody array and treated as in F. Values are the mean of biological duplicates and are represented as ratio to DMSO-treated control cells. Data in A–D and F–I are presented as mean ± SEM of biological replicates. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.