Figure 5. SLCA71 promoter activity in response to insulin.

(a) Luciferase (Luc) reporter constructs containing serial truncations of SLC7A1 promoter (−1606 bp (pGL3-hCAT-1⁻¹⁶⁰⁶) or −650 bp (pGL3-hCAT-1⁻¹⁶⁰⁶) from the transcriptional start point) were transfected in primary cultures of HUVECs incubated (8 hours) without (plain bars) or with (dashed bars) 10 μmol/L nitrobenzylthioinosine (NBTI). Cell transfection was done in the absence or presence of 1 nmol/L insulin and/or ZM-241385 (10 nmol/L), along with Renilla reporter plasmid, and assayed for Firefly and Renilla luciferase activity, respectively. Results depict ratio of Firefly/Renilla luciferase activity. Cells were also transfected with the empty pGL3-basic vector or pGL3-control vector (SV40 pGL3) as negative or positive controls, respectively. (b) Fraction of SLC7A1 reporter constructs activity inhibited (sensitive) by ZM-241385 in absence (Basal) or presence of insulin or NBTI as in (a). (c) SLC7A1 reporter constructs fraction activity non-inhibited (insensitive) by ZM-241385 as in (a). In (a), *P<0.05 versus Control, †P<0.05 versus corresponding values in the presence of ZM-241385. In (b), *P<0.05 versus corresponding Basal, †P<0.05 versus values in pGL3-hCAT-1⁻¹⁶⁰⁶ in the presence of insulin, ‡P<0.05 versus values in pGL3-hCAT-1⁻⁶⁵⁰ in the presence of insulin or insulin + NBTI. Values are mean ± SEM (n = 6).