Figure 3
From: Myc interacts with Max and Miz1 to repress C/EBPδ promoter activity and gene expression
Miz1 functions in Myc repression of C/EBPδ promoter activity. A. HC11 cells were co-transfected with Miz1 or a vector control plus a C/EBPδ promoter luciferase construct and Renilla control. Luciferase activity was assessed under Growing (Gr) (Myc expressed, C/EBPδ repressed) or growth arrest (GA) (Myc not expressed, C/EBPδ expressing) conditions. Luciferase results were normalized to the Renilla control. C/EBPδ promoter driven luciferase results from Miz1 transfected cells are expressed relative to the vector control, "Gr" results which were set as "1". B. Lysates from luciferase assays (A.) were analyzed by Western blot to assess Myc and Miz1 protein levels. β-actin levels were assessed as a loading control. C. HC11 cells were transfected with Miz1 siRNA treatment and C/EBPδ promoter driven luciferase assays performed as described. Luciferase results were normalized to the Renilla control. C/EBPδ promoter driven luciferase results from Miz1 siRNA treated cells are expressed relative to the vector control, "Gr" results which were set as "1". D. Lysates from luciferase assays (C.) were analyzed by Western blot to assess Myc and Miz1 protein levels. β-actin levels were assessed as a loading control. Luciferase results for the Miz1 expression and Miz1 siRNA treatment groups shown are the average-fold changes relative to the "scrambled" siRNA values from 2 independent experiments with duplicates performed in each experiment. E. HEPG2 cells were co-transfected with Miz1 or a vector control plus a low density lipoprotein receptor (LDLR) promoter luciferase construct and Renilla control. Luciferase results were normalized to the Renilla control. LDLR promoter driven luciferase results from Miz1 transfected cells are expressed relative to the vector control results which were set as "1".