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. 2008 Jun 16;105(24):8440–8445. doi: 10.1073/pnas.0803096105

Fig. 2.

Fig. 2.

Effect of BPIPP on cAMP accumulation in cultured cells. (A) RFL-6 cells were pretreated with 50 μM BPIPP or vehicle for 10 min and then with 50 μM forskolin (For) or 100 μM isoproterenol (Iso); cells were pretreated with 1 μg/ml cholera toxin for 30 min (CT30) or 60 min (CT60) and then, after a 10-min incubation with IBMX and BPIPP, cAMP accumulation was measured. *, P < 0.01; n = 4. (B) T84 cells were pretreated in serum-free growth medium in the presence of 1 μg/ml adenylyl cyclase toxin from Bordetella pertussis (BAC), a combination of 2 μg/ml protective antigen and 0.1 μg/ml edema factor from Bacillus anthracis (edema toxin; ET), or 1 μg/ml Vibrio cholera toxin (CT) for 1 h (pretreatment phase; pretreat); cells were washed and further incubated in DBPS containing 1 mM IBMX for 10 min to measure cAMP accumulation (incubation phase; incubate); cells were treated with vehicle or with 50 μM BPIPP during the corresponding phase as indicated. Control (100% values) for BAC, ET, and CT were 9.98 ± 0.72, 0.14 ± 0.01, and 7.85 ± 0.82 nmol of cAMP per mg of protein. *, P < 0.05; **, P < 0.01; n = 3. (C) T84 cells were pretreated with 50 μM BPIPP or vehicle and 1 mM IBMX for 10 min and then treated with indicated concentrations of forskolin for 10 min. n = 4.