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Figure 2. Regulation of adenosine receptor engagement by ecto-ADA. (A) Regulation of cAMP responses by ecto-ADA. Human gingival fibroblasts (5.0 x 105 cells/well) were incubated with or without Jurkat cell lysate for 30 min at 37°C and stimulated with 20 µM adenosine for 5 min at 37°C in the presence or absence of 5 µM dCF. The data shown are the mean ± SD of 4 cAMP determinations and are representative of 3 separate experiments. *p < 0.05 compared with the lysate (–) dCF (–) group. **p < 0.05 compared with the lysate (+) dCF (–) group. (B) Effects of adenosine (Ado) on HAS mRNA expression in human gingival fibroblasts. Human gingival fibroblasts were cultured with or without Ado (20 µM) for 3 hrs, and then RT-PCR was carried out for the detection of HAS1, HAS2, and HAS3 mRNA expression. The results shown are representative of 3 separate experiments. The number of PCR cycles is shown above each lane. Each band was standardized against the amount of HPRT. (C) Effects of ecto-ADA on adenosine (Ado)-induced HAS1 mRNA expression in human gingival fibroblasts. Human gingival fibroblasts were pre-incubated with or without Jurkat cell lysate for 30 min at 37°C, and were treated with Ado (20 µM) with or without dCF (5 µM) for 2.5 hrs. HAS1 mRNA expression was examined by RT-PCR. Results of 1 representative experiment from among 3 identical experiments are shown. The number of PCR cycles is shown above each lane. Each band was standardized against the amount of HPRT.
