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Figure 2 | Molecular Cancer

Figure 2

From: Myc interacts with Max and Miz1 to repress C/EBPδ promoter activity and gene expression

Figure 2

Myc interacts with Miz1; Miz1 plays a key role in Myc repression of C/EBPδ promoter activity. A. HC11 cell lysates were immunoprecipitated with anti-Myc and anti-Miz1 antibodies and the immunoprecipitates analyzed by Western blot using anti-Myc, Miz1 and Sp1 antibodies. "Input": western blot analysis of HC11 whole cell lysates (positive control). "IgG": nonspecific rabbit IgG immunoprecipitates (negative control). B. HC11 cell chromatin was immunoprecipitated using antibodies against Myc or Miz1. Immunoprecipitated DNA was amplified using primers flanking the C/EBPδ proximal (P200) promoter region and the distal (P1.8K) C/EBPδ upstream promoter regions. "Input" results are derived from direct PCR amplification of P200 and P1.8K C/EBPδ promoter regions from HC11 genomic DNA (Positive control). IgG: nonspecific rabbit IgG immunoprecipitated (negative control). C. HC11 cells were transfected with vector control, Myc wt or Myc V394D expression constructs (V5-tagged). Co-immunoprecipitation was performed using an anti-V5 antibody. Immunoprecipitates were analyzed by Western blot using anti-Miz1 and anti-V5 antibodies. D. HC11 cells were co-transfected with a C/EBPδ promoter luciferase reporter construct, vector control, Myc wild type (wt) or Myc V394D. C/EBPδ promoter driven luciferase activities were normalized to renilla luciferase activity. Results for the Myc transfected cells are expressed relative to the vector control results which were set as "1". E. Whole cell lysates (20 ul) from lucifease assays in (D.) were analyzed by Western blot to assess Myc wt and Myc V394D expression. Luciferase results shown are the average-fold changes relative to the vector control values from 2 independent experiments with duplicates performed in each experiment.

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