Figure 4.

Regulatory relationship of miR-429 to PD-L1. (A) Cells were transfected with mimic-429 and PD-L1 luciferase reporter plasmids [wild type (WT) and mutant (MUT)] and dual-luciferase reporter assays were performed. (B) Luciferase reporter plasmids (WT and MUT) of inhibitor-429 and PD-L1 were transfected into HGC27 cells. Luciferase activity was assessed via dual-luciferase reporter assays. (C) Mimic-429 was transfected into SGC7901 and BGC823 cells and the expression of PD-L1 was detected by quantitative PCR (qPCR). (D) Mimic-429 was transfected into SGC7901 and BGC823 cells and the expression of PD-L1 was assessed via Western blot. (E) Inhibitor-429 was transfected into MKN45 and HGC27 cells and the mRNA expression of PD-L1 was assessed by qPCR. (F) Cells were transfected with inhibitor-429 and the expression of PD-L1 was assessed by Western blot. (G) Mimic-429-expressing BGC823 cells were assessed for PD-L1 localization and expression by immunofluorescence analysis. Scale bar, 50 μm. (H) Inhibitor-429 was transfected into HGC27 cells and the intracellular localization and expression of PD-L1 were assessed by immunofluorescence analysis. Scale bar, 50 μm. Results are the mean ± SEM of triplicate samples from three independent experiments. *P < 0.05.