VHL Frameshift Mutation as Target of Nonsense-Mediated mRNA Decay in Drosophila melanogaster and Human HEK293 Cell Line
Figure 3
Effect of CHX treatment on the level of the c.163delG and c.239delG VHL mRNA. (a) Scheme of the WT (upper panel) and c.163delG constructs, which contained the exons 1-2 (marked in the boxes by numbers), a part of the intronic sequence between exons 1-2 and a piece of intron downstream the exon 2. The CMV promoter is marked by a thick horizontal arrow. (b) Scheme of the WT and c.239delG construct containing the entire coding sequence of the VHL gene. (c) and (d) QPCR analysis of VHL transcripts before and following CHX treatment. The level of mRNA transcribed from VHL construct carrying either the wild type sequence or the c.163delG and c.239delG mutations was normalized to the mRNA level of GFP. The ratio between these normalized levels following CHX treatment was calculated and compared with the ratio in untreated cells. The fold increase in the level of VHL c.163delG and c.239delG transcripts is shown as mean SEM.
