c-MYC promoter activity induced by GSE24-2. (A) Schematic representation of the c-MYC promoter indicating the different constructs used in the experiments. (B) 293T pLNCX and 292T GSE24-2 cells were transfected with the different constructs of the c-MYC-luc reporter (1 μg per million cells). Luciferase activity was measured following 24 hours of transfection. CMV-Renilla (0.1 μg/mL per million cells) was used as a control for transfection efficiency. Data points represent the mean and standard deviations of 2 experiments performed in quadruplicate. (C) Schematic representation of the mutations generated in NHEIII. (D) The indicated cell lines were transfected with the mutants of the px3.2 reporter (1 μg per million cells). Luciferase activity was measured following 24 hours of transfection. CMV-Renilla (0.1 μg/mL per million cells) was used as a control for transfection efficiency. Data points represent the mean and standard deviations of 2 experiments performed in quadruplicate. (E) MEFs were transfected with pLNCX-, DKC-, or GSE24-2–expressing vectors. Following 24 hours of transfection, total RNA was isolated and expression of mouse c-myc and β-actin RNA was estimated by RT-PCR. The experiment was repeated 3 times, with similar results. (F) MEFs were transfected with pLNCX-, DKC-, or GSE24-2–expressing vectors. Following 24 hours of transfection, total protein extracts were obtained and expression of mouse c-myc and α-tubulin were detected by immunoblot using specific antibodies. The experiments were repeated 3 times, with similar results.