Figure 6.

Knockdown of gga-miR-375 expression promotes the cell cycle and cell proliferation by targeting YAP1. (A) DF-1 cells transfected with anti-gga-miR-375 had decreased gga-miR-375 mRNA expression at 48 h as assessed by reverse transcription-quantitative PCR. Anti-gga-miR-con and mock groups are the controls. (B) Protein expression of YAP1 was measure after DF-1 cells were transfected for 48 h with anti-gga-miR-375. (C) Protein expression levels of YAP1 after 48 h transfection with anti-gga-miR-375 or co-transfected with anti-gga-miR-375 + siRNA-YAP1. (D) Knockdown of gga-miR-375 promoted cell proliferation by targeting YAP1, as identified by Cell Counting Kit-8 assay results after 24, 48 and 72 h. (E) Flow cytometry results indicated that knockdown of gga-miR-375 expression promoted the cell cycle by targeting YAP1. The cell cycle assay was performed after knockdown of gga-miR-375 or gga-miR-375 + YAP1 in DF-1 cells and evaluated with a FACSCalibur flow cytometer 48 h after transfection. (F) DF-1 cells transfected with anti-gga-miR-375 or anti-gga-miR-375 + siRNA-YAP1 were harvested 48 h after transfection for western blot analysis with antibodies against YAP1, cyclin D1, cyclin E and β-actin. Data are presented as the mean ± SD of three independent experiments. ^**P<0.01. YAP1, yes-associated protein 1; miR, microRNA; siRNA, small interfering RNA; PI, propidium iodide; OD, optical density; Scra, scramble.