Fig. 5.

VEGF restored the down-regulated eNOS mRNA and protein levels by the intronic small RNA. HAECs were transfected with indicated RNA duplex. After 12-h transfection, 10 ng/ml of VEGF was added into the media. (A) Total RNA was collected after 24-h transfection for the analysis of mRNA levels by the real-time PCR. Columns represent mean + SEM of three independent experiments (*, P < 0.05 vs. non-VEGF; **, P < 0.01 vs. control by Student's t tests). (B) Protein samples were collected after 48-h transfection and measured by Western blotting. Lanes 1, 2, 3, and 4 represent control (19-nt siRNA targeting luciferase gene), 19-nt exonic siRNA, 19-nt intronic siRNA, and 27-nt intronic microRNA at the dose of 50 nmol/liter, respectively.