Figure 2. MCT-1 functions as a receptor for ancestral HERV-T.
(A) Scheme of the receptor screening strategy. DF-1 cells were transduced with a lentiviral cDNA library. Two days later, the cells were challenged with MLV-ancHTenv containing a neo gene. After a further two days, cells were placed in G418 selection. After another 10 days, G418-resistant cells were replated and challenged with MLV-ancHTenv containing a hygromycin resistance gene. Two days later cells were placed in hygromycin selection. After a further 10 days, Hygromycin-resistant cells were replated and challenged with MLV-ancHTenv containing RFP and were found to be highly susceptible to infection (Figure 2—figure supplement 1A). Genomic DNA (gDNA) was extracted from this cell population and subjected to PCR using primers specific to the lentiviral vector (Figure 2—figure supplement 1B). (B) Fluorescent micrographs of 293T and DF-1 cells, expressing hMCT1 or a control protein, following infection with MLV-ancHTenv expressing GFP as reporter. Scale bar = 200 μm. (C) Titers of MLV-ancHTenv/GFP on 293T and DF-1 cells expressing hMCT1 or a control protein (Mean ± SD, n = 2 replicates, one of two separate experiments).
Figure 2—figure supplement 1. HERV-T receptor identification.
