Fig. 4.

SAE1 knockdown induced G2 phase arrest and apoptosis of glioma cells. a SAE1 knockdown induced G2 phase arrest of glioma cells. The specific SAE1 siRNA3 was transfected into U87 and U251 cells for 48 h, following cells were staining with propidium idodide (PI) and assayed by flow cytometry. Quantification of cell cycle distribution was shown in the right panel. Data were presented as mean ± SD of three separate experiments. b SAE1 silencing induced apoptosis of U87 and U251 cells. Cell apoptosis was detected by Annexin V-PI double staining and flow cytometry analysis. The proportion of early apoptotic cells (annexin V positive) and late apoptotic cells (Annexin V and PI positive cells) were shown. Data were presented as mean ± SD of three separate experiments. *p < 0.05. c The expression of SAE1, AKT signaling pathway proteins, typical cell cycle-related proteins (CDK2, Cyclin D1, p21) and typical apoptosis-related proteins (Bcl-2, active Caspase-3) were detected by Western blot after siSAE1 treatment for 48 h. Each has the expression of β-actin as internal control. siCon: non-targeting siRNA. siSAE1: The SAE1 siRNA3 that specifically inhibits SAE1. d The AKT signaling pathway proteins and several typical cell cycle or apoptosis-related proteins were changed due to SAE1 overexpression. The β-actin was taken as internal control. Con: pFlag empty vector as a mock control. SAE1: pFlag-SAE1 transfection