Fig. 2.
Characterization of siRNA for A2A. HUVECs were isolated from normal pregnancies and exposed to siRNA for A2A (siRNA-A2A) for 48 h as described in Methods section. a Representative image of semiquantitative PCR assays for A2A (A2A) and their respective control loading β-actin. Positive control (C+, HUVEC without transfection). Lanes 1, 2, and 3 show different doses used in RNA interference (0.3 to 1 μg/μl). b HUVEC immunocytochemistry for the presence of protein A2A, in the absence (Control) or presence of siRNA-A2A. c Testing of survival determined by cell counting with trypan blue in the absence (−) or presence (+) of siRNA-A2A. d Quantification of intracellular cAMP in HUVEC cells in the absence (−, white bar) or presence (+, hatched bar) of siRNA-A2A and CGS-21680 (10−8 M). e Quantification of nitrite in culture medium as described in d. The results are expressed as medians and interquartile ranges. In b, black bar represents 70 μm. d *p < 0.05 vs respective control in the absence of CGS-21680. In e, *p < 0.05 compared with control without CGS-21680 or siRNA for A2A. †p < 0.05 vs CGS-21680 in the absence siRNA for A2A. n = 3–5 for each assay in duplicate