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. Author manuscript; available in PMC: 2009 Dec 29.
Published in final edited form as: Ann N Y Acad Sci. 2005 May;1041:296–299. doi: 10.1196/annals.1282.044

FIGURE 2.

FIGURE 2

Overexpression of ACV but not ACII in MCF-7 cells leads to relaxin stimulation of cAMP. MCF-7 cells (4 × 105 cells per treatment) were transfected with 0.3 μg pcDNA3-ACII (A) or 0.3 μg pcDNA3-ACV (B) for 36 h using Fugene 6 transfectant reagent and starved overnight in medium without fetal bovine serum. Transfected cells were pretreated in the presence of phosphodiesterase inhibitor (500 μM IBMX) for 15 minutes and then treated with vehicle, 0.1 μg/ml relaxin (rlx), 0.1 μg/ml relaxin + 30 nM forskolin (Fsk), or 30 nM forskolin for 20 min. Treatments were stopped by removal of media and addition of 0.1 N HCl. Intracellular cAMP was detected by enzyme immunoassay. (A) Data are expressed as the mean ± SE from a single experiment and are representative of two different experiments. (B) Data are expressed as the mean ± SE from three different experiments each performed in duplicate. Significant differences (*P <.05) between groups are designated (t test).