Knockdown of NHERF-2 increases endothelial proliferation. (A) Only NHERF-2 was expressed in HUVECs. The breast cancer cell line MCF-7 expressed both NHERF-1 and NHERF-2. (B) HUVECs were transfected with control-scrambled or NHERF-2 siRNA for 48 hours. The cells were starved overnight and stimulated with 10 ng/mL VEGF for 24 hours, followed by [³H]-titrated thymidine incorporation assay represented here in fold with respect to the control-scrambled unstimulated cells as in C-V. (C) Approximately 5-fold higher expression of NHERF-2 in HAEND cells compared with HSECs (left). Efficient knockdown of NHERF-2 in HAEND cells as determined by real-time PCR (middle). HAEND cells were transfected with control-scrambled or NHERF-2 siRNA for 48 hours, followed by [³H]-titrated thymidine incorporation assay (right). Statistical significance was determined with 2-sided Student t test, *P < .05 was considered significant.