Figure 6

Cyclin D1 expression is reduced in CNE1-LMP1 cells after treatment with EGFR siRNA and STAT3 siRNA. (A) Dual luciferase-reporter assays were performed in CNE1-LMP1 cells after co-transfection with either control siRNA (siControl), EGFR siRNA (siEGFR), or STAT3 siRNA (siSTAT3) in addition to cyclin D1 promoter-reporter constructs and a Renilla luciferase transfection control plasmid. Firefly luciferase was measured and normalized to Renilla luciferase activity. The fold change in cyclin D1 expression by the indicated siRNA is displayed in each case. The control siRNA served as a non-targeting control. (mean ± SD, n =3, *p < 0.05) (B) The cells were incubated with medium containing the indicated siRNAs for 72 h. Total RNA was isolated from the cells and subjected to real-time PCR, using specific primers designed to amplify cyclin D1. β-actin mRNA served as an internal control. (mean ± SD, n =3, *p < 0.05, **p < 0.01). (C) Western Blot was performed in CNE1-LMP1 cells after co-transfection with the indicated siRNAs for 72 h. β-actin was served as an internal control. (D) FACS was performed in CNE1 and CNE1-LMP1 cells after co-transfection with the indicated siRNAs for 72 h. The data are presented from three independent experiments.