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. 2020 Mar 12;12(3):666. doi: 10.3390/cancers12030666

Figure 2.

Figure 2

Effect of pemetrexed on PD-L1 expression in A549 cell line. (A) A549 cells were treated with 100 nM pemetrexed for the indicated period of time and PD-L1 mRNA level, evaluated by RT-PCR, was reported. (B) Time-dependent modulation (100 nM pemetrexed) and (C) dose-dependent modulation (72 h) of PD-L1 protein expression in A549 cells were evaluated by western blotting. A549 cells were continuously exposed to 500 nM pemetrexed for the indicated period of time or treated for 24 h and, after drug removal, the cells were incubated with fresh medium for 24 h or 48 h. At the indicated times, total PD-L1 protein, membrane PD-L1 protein, and PD-L1 mRNA were quantified by western blotting (D), flow cytometry (E), and RT-PCR (F), respectively. * p < 0.05; ** p < 0.01; *** p < 0.001. Data in (A), (E), and (F) are mean values ± SD of three independent experiments. Results in (B–D) are representative of three independent experiments.