Figure 2.

TAAR1 induces CREB phosphorylation and Irs-2 gene expression in Ins-1 cells. A, Western blots of CREB phosphorylation in response to T₁AM (10 μm) and forskolin (0.3 μm) for 5–30 min in the presence (11 mm) or absence of glucose. B–D, addition of 10 μm MDL-12,330A (B), 10 μm H89 (C), or 150 nm PKA Cat-α siRNA (D) inhibits CREB phosphorylation (10 min) induced by T₁AM and forskolin. Representative blots of pCREB from one of at least three independent experiments are shown; blots were stripped and reprobed with β-actin as a loading control. E, quantitative PCR of Irs-2 gene expression (1 h) induced by 10 μm T₁AM in the presence and absence of MDL-12,330A (10 μm) or LY294002 (10 μm, inhibits PI3K) pretreatment. Data are expressed as mean ΔΔCT (± S.D.) of Irs-2 using Gapdh as the housekeeping gene and were analyzed by one-way ANOVA (global p values are shown) using Dunnett's multiple comparison test, comparing all columns with 10 μm T₁AM treatment (n = 3). ***, p < 0.001.