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. 2010 Jun 9;151(8):3589–3599. doi: 10.1210/en.2010-0106

Figure 7.

Figure 7

EGR1 regulates CART promoter activity. A, In cultured vagal afferent neurons transfected with 3.45 kb CART-luc promoter-reporter construct, incubation with 10 nm CCK8s (16 h) increased luciferase activity compared with unstimulated cells. Mutation of a putative EGR1 binding site in the CART-luc (CARTΔEGR1) vector reduced the response to CCK, and so too did pretreatment with EGR1 siRNA (***, P < 0.001 compared with control; *, P < 0.05). B, Cotransfection of CART-luc with an EGR1 expression vector (EGR1OE) dramatically increased the response to CCK (***, P < 0.001) and also increased basal expression; however, when the EGR1 expression vector was cotransfected with CARTΔEGR1, there was no change in basal expression and no additional response to CCK. C, Concentration-response relationships for CART-luc with graded concentrations of CCK8s alone and in the presence of 10 ng/ml leptin showing potentiating between CCK8s and leptin, which together stimulated approximately 7-fold increases in maximal response and induced significant responses at 10 pm CCK8s. D, The synergistic effects of leptin on CCK-induced CART-luc were inhibited by the STAT3 inhibitor AG490 and by Ro32-0432 and U0126 (*, P < 0.05 compared with CCK8s and leptin).