Figure 5
From: PTEN inhibits BMI1 function independently of its phosphatase activity
PTEN reduces hTERT promoter activity via inhibiting BMI1 function. DU145 cells were transfected with 100 nM PTEN siRNA, BMI1 siRNA, PTEN siRNA plus BMI1 siRNA, and the respective control (Ctrl) siRNA (Dharmcon) using LipofectAMINE2000 (Invitrogene) for 48 hours following our published procedure [52]. The expression of individual proteins was examined by western blot using specific antibodies (top panel). These cells were transfected with a pGL3-hTERTmin-Luc reporter, a lacZ vector, and plus the indicated siRNAs for 48 hours. Luciferase activities were determined and normalized against the respective lacZ activity. Experiments were carried out in triplicate and were repeated three times. Average data of these independent experiments is shown. Luciferase activities in PTEN siRNA (p < 0.001) and BMI1 siRNA (p < 0.05) cells are significantly different from that in Ctrl siRNA cells (bottom panel).