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. 2021 Sep 7;112(11):4457–4469. doi: 10.1111/cas.15110

FIGURE 3.

FIGURE 3

Chemokine C‐C motif ligand 21 (CCL21) upregulated programmed death‐ligand 1 (PD‐L1) expression to promote immune escape in vitro. A, Western blot analysis of PD‐L1 abundance in MIA‐PaCa‐2 and SW1990 cells treated with CCL21 at the concentration of 0, 12, 25, 50, or 100 ng/mL for 48 h. B, Western blot analysis of PD‐L1 abundance in MIA‐PaCa‐2 and SW1990 cells treated with CCL21 at a concentration of 50 ng/mL for 0, 12, 24, 36, and 48 h after 24 h starvation. C, Flow cytometry analysis of PD‐L1 expression in MIA‐PaCa‐2 and SW1990 cells treated with 50 ng/mL CCL21 for 48 h. D, Quantitative real‐time PCR analysis of PD‐L1 mRNA level when MIA‐PaCa‐2 and SW1990 cells were treated with CCL21 (50 ng/mL) or PBS for 12 h. E, T cell‐mediated tumor cell killing assay analysis of different groups of cancer cell survival after coculture with or without activated T cells or anti‐PD‐L1 Ab for 4 d. F, Modified T cell‐mediated tumor cell killing assay using the Transwell system. Pancreatic cancer cells were seeded in the lower chamber prior to addition of the corresponding tumor cell line‐specific T cells in the upper chamber. Surviving tumor cells were visualized by crystal violet staining