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. 2020 Jul 10;8:540. doi: 10.3389/fcell.2020.00540

FIGURE 5.

FIGURE 5

MiR-138-5p targets PD-L1 to decrease proliferation A549 cells. (A) Diagram of the has-miR-138-5p binding site in wild-type (WT) human or mouse PD-L1 3′UTR. The mutation was generated at the binding site (Mut). (B) In a dual-luciferase reporter assay, a luciferase reporter gene containing the WT or Mut human or mouse PD-L1 3′UTR was co-transfected with NC or miR-138-5p mimic into HEK293T cells, and relative luciferase activities were assessed. The gene (C) and protein expression (D) of PD-L1 on A549 cells/3LL cells, which were transfected with NC or miR138 mimic or not. (E) Upon NC, shRNA PD-L1 treatment, the expression levels of CCND3, CDC20, MCM2, and Ki67 were assessed by RT-QPCR. After NC or shRNA PD-L1 silencing in A549 cells, cell proliferation was analyzed by flow cytometry detection of Ki67 (F) or the MTT assay (G). (H) Percentages of apoptotic A549 tumor cells co-cultured with DCs, which were pretreated or not with shRNA PD-L1 or NC. (*P < 0.05, **P < 0.01, ***P < 0.001).