Effect of β3AR agonist stimulation on phosphorylation of CREB, ATF-1, and ATF-2. 3T3-L1 cells were stimulated with CL316243 (1 μM) for 3, 30, and 60 min, then cell lysates were subjected to Western blot analyses with antibodies to pCREB, pATF-1, and pATF-2. CL316243 stimulation increased phosphorylation of pCREB, pATF-1, and pATF-2 (a). 3T3-L1 cells were pretreated with SB203580 (10 μM for 1h) or RO31-8220 (10 μM for 1 h) before stimulation with CL316243 (1 μM for 45 min). CL316243-induced phosphorylation of CREB and ATF-2 was attenuated in the presence of SB203580, while ATF-2 phosphorylation was also decreased in the presence of RO31-8220, although non-specific phosphorylation of ATF-2 also has been observed in presence of inhibitors alone. ATF-1 phosphorylation was not affected by inhibitor treatment. β-actin was used as the loading control (b).