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. 2022 May 9;7(9):e152731. doi: 10.1172/jci.insight.152731

Figure 5. Targeting MNKs does not affect PD-L1 expression.

Figure 5

(A and B) Mouse pancreatic (KPC-344) and thyroid (TBP-3868) cancer cells were pretreated with DMSO, CGP57380 (CGP; 1, 10 μM), or eFT508 (eFT; 0.01, 0.05, 0.1 μM) for 24 hours and then treated with either vehicle control or IFN-γ (200 ng/mL) for additional 24 hours. The cells were then analyzed for PD-L1 expression by flow cytometry, and the mean fluorescence intensity (MFI) was calculated using FlowJo. (C) Mouse cancer cells were transfected with siRNAs targeting Mknk1 and Mknk2 (siMNK1+2) for 48 hours. Knockdown efficiency was confirmed by Western blotting for MNK1 and qPCR for Mknk1 and Mknk2 mRNAs. Data are shown as the mean ± SD from 2 technical replicates. Data are representative of 3 independent biological replicates. The cells were then analyzed for PD-L1 by flow cytometry, and the MFI was calculated using FlowJo. In C, data are shown as the mean ± SEM from 3 biological replicates. Unpaired t test. In A and B, data are shown as the mean ± SEM from 3 biological replicates, and analysis was performed by 2-way ANOVA; ***P < 0.001; ****P < 0.0001.