Figure 3.

Different c-Myb-mediated synergy on natural chromatin-embedded promoters. (A) Schematic presentation of the regulatory elements of the mim-1 and lysozyme genes according to (35–37). Both genes contain functional MREs in enhancer elements as well as in promoters. Question mark indicates functionality uncertain. HD11 cells were transfected with plasmids expressing c-Myb-HA or the sumoylation-negative mutant c-Myb-2KR-HA. (B) A western blot performed with anti-HA antibody, using lysates from the same cultures used for activity measurements. The α-tubulin was used as loading control. Activation of the endogenous Myb-target genes (C) mim-1 and (D) lysozyme was measured by quantitative real-time PCR. Target gene expression data were normalized by the relative expression of the housekeeping gene HPRT1 and represented as relative to empty vector-transfected cells, which were set to 100. The results represent the mean ± SEM of three independent biological assays, each analysed in duplicate for expression levels.