Davis et al. BMC Cancer 2011, 11:173 http://www.biomedcentral.com/1471-2407/11/173 Page 5 of 7 Figure 2 Aberrant promoter hypermethylation is rare in the MAP2K4 promoter region. A. Map of MAP2K4 promoter region from UCSC Genome browser (Mar. 2006 build) showing the gene in blue (exon 1 indicated by thicker bar), predicted CpG island in green, location of all CpG dinucleotides in orange, and location of the primers and MS-SSCP PCR product in black. MS-SSCP (B) and DNA sequencing electrophoretograms (C) for a representative tumor sample lacking methylation (top) and the positive methylation control SssI treated normal lymphoblast DNA. Figure 3 Loss of MAP2K4 expression affects cellular proliferation. Cellular proliferation was assessed using alamarBlue dye measured by fluorescence at 540/610 nm. Each of three experiments was performed in triplicate and normalized to the 24 h time-point. The average of three biological replicates is shown, except for the 168 h time-point, which was only done twice. Statistical significance was determined by 2-way ANOVA. Error bars are representative of SEM, n = 3.