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. 2021 Nov 1;131(21):e137750. doi: 10.1172/JCI137750

Figure 1. Pulmonary melanoma metastasis stimulates PD-L1.

Figure 1

Eight-week-old WT mice were challenged with B16-F10 (B16) melanoma cells or control vehicle (PBS) via tail vein injection and evaluated 2 weeks later. (A) Real-time reverse transcriptase (RT) PCR was used to quantitate the levels of mRNA encoding PD-L1 in the lungs from mice treated i.v. with PBS (B16 –) or B16 cells (B16 +). Each dot represents an evaluation in an individual animal. (B) Western blot evaluations of PD-L1 accumulation in lungs from mice treated with PBS (B16 –) or B16 cells (B16 +). (C and D) FACS evaluations quantitating the accumulation of PD-L1 in cell populations from lungs of mice treated with B16 cells (B16-F10 +) or vehicle control (B16-F10 –). These evaluations used specific markers of airway epithelial cells (CC10), alveolar epithelial cells (surfactant apoprotein C [SP-C]), T cells (CD3), and macrophages (CD68). (E) Representative double-label fluorescent immunohistochemical evaluations in lungs from mice challenged with B16 melanoma cells using cell-specific markers (alveolar epithelial cells, SP-C; macrophages, CD68) (red) and anti–PD-L1 (green). The arrows highlight cells that stained with both antibodies. The values in A represent the mean ± SEM of the noted evaluations represented by the individual dots. B–E are representative of a minimum of 2 similar evaluations. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s t test). Scale bars: 50 μm.