Figure 2.

Overexpression of LATS2 suppresses cell proliferation by delaying G₁/S transition in U-372 MG cells. (A) Efficiency of LATS2 plasmid and siLATS2 transfection was validated in U-372 MG cells. The mRNA and protein expression levels of LATS2 were significantly upregulated or downregulated in cells transfected with plasmids or siRNA, respectively. Empty vector or NC siRNA were used as controls. (B) Effects of LATS2 on cell growth were determined by Cell Counting Kit-8 assay. OD₄₅₀ was measured at 0, 24, 48 and 72 h post-transfection. Overexpression of LATS2 decreased cell growth (left), whereas LATS2 knockdown increased cell growth (right). (C) Effects of LATS2 on cell cycle progression were measured by flow cytometry using propidium iodide. Overexpression of LATS2 increased the percentage of cells in the G₁ phase and decreased the number of cells in the S phase compared with the control, whereas knockdown of LATS2 had the opposite effects; G₂/M phase was not affected. (D) Protein expression levels of cyclin D, CDK4 and CDK6 were determined by western blotting and β-actin was used as an internal reference. Expression levels of these proteins were decreased in LATS2-overexpressed cells and were increased in LATS2 knockdown cells. All data are presented as the means ± standard deviation (n=3). *P<0.05; **P<0.01; ***P<0.001 compared with control or siNC groups. CDK, cyclin-dependent kinase; LATS2, large tumor suppressor kinase 2; OD, optical density; NC, negative control; si/siRNA, small interfering RNA.