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Fig. 4 | Molecular Medicine

Fig. 4

From: The Regulation and Localization of Angiopoietin-1, -2, and Their Receptor Tie2 in Normal and Pathologic Human Placentae

Fig. 4

Effects of reduced oxygen on Ang-1 and Ang-2 mRNA levels. Effects in the (A) first trimester (n = 3) and (B) term villi cultures (n = 3) detected by semiquantitative RT-PCR for Ang-1 and Ang-2. cDNA samples were normalized by RT-PCR with primer set detected β-actin. The proper numbers of cycle for each RT-PCR reaction were determined in a pilot study in which different numbers of amplification cycles were performed, for Ang-1 and Ang-2: 28, 30, 32, 35, 38, and 40; for β-actin: 18, 20, 25, 30. The optimal number for PCR amplification cycles was determined and chosen in the mid-exponential phase. Subsequent PCR amplification on β-actin matched samples for Ang-1 and Ang-2 were carried out. The reaction products were quantified by inclusion of a small amount of 32P-labeled CTP (0.01 µCi in total 20 µl reaction) in each reaction. Autoradiography and denstometric quantitation of the signals were then carried out. The entire procedure was repeated three times. Data are presented as mean ± SEM. Statistical comparisons were for paired data under ambient (open column) and reduced oxygen (solid column) conditions. *p < 0.05. Lanes 1–3, ambient oxygen; lanes 4–6, reduced oxygen.

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