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. 2016 Sep 18;7(44):71255–71273. doi: 10.18632/oncotarget.12104

Figure 9. GILT decreases the infectivity of released MLV particles.

Figure 9

A. COS7 cells were transfected with the MLV vector construction plasmids, together with pcDNA3.1 or wild type GILT. The culture supernatants of the transfected cells were used to inoculate TE671 cells. Relative values to titers in pcDNA3.1-transfected cells are indicated (n = 3). Asterisks indicate statistically significant differences. B. COS7 cells were transfected with the MLV vector construction plasmids, together with pcDNA3.1, wild type GILT, or the DCS mutant. Cell lysates and virion pellets from the transfected cells were analyzed by western blotting, using anti-MLV p30 and anti-GILT antibodies. C. COS7 cells were transfected with the MLV vector construction plasmids, together with pcDNA3.1, the C-terminally HA-tagged wild type CD63, or the DCS or TCS mutant. Cell lysates and virion pellets from the transfected cells were analyzed by western blotting, using anti-MLV Gag, anti-HA, and anti-actin antibodies. D. COS7 cells were transfected with the MLV vector construction plasmids, together with pcDNA3.1 or wild type GILT. Cell lysates and virion pellets from the transfected cells were analyzed by western blotting, using anti-MLV p30 (CA), anti-SU, anti-TM, anti-GILT, and anti-actin antibodies. The C-terminal R peptide of the MLV TM protein is cleaved after virion budding. Therefore, the R peptide-containing TM (TM+R) and the R peptide-deficient TM (TM-R) proteins were detected in cell lysates and virion pellets, respectively.