Figure 2.
Engineered SVA retrotransposition is mediated by human L1 proteins in HeLa HA cells. Different marked SVAs, ORF1 mneoI and Alu neoTet were co-transfected with various drivers (A–G) to determine the role of L1 proteins in SVA retrotransposition. All transfections were carried out in six-well plates with 1.5 µg of the corresponding ‘driver' plasmid and 0.5 µg of the corresponding ‘passenger' plasmid. Data are presented as the mean number of G418R foci per well ± SEM, with the number of replicates (n) below each mean. Where no data are presented, it means that the experiment was not carried out. ‘Hot' L1s, L1-RP and L1.3 mobilize engineered SVAs (A and B). Removal of the CMV and L1 promoter (5′ UTR) from pcDNA.L1-RP reduces SVA foci formation to background levels (C). Different drivers containing point mutations (D and G) or lacking ORF1 coding sequence (E and F) were co-transfected with SVA. (*) indicates the relative location of the engineered point mutation.