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Table 1 Infection assay on human cells.

From: The effects of N-terminal insertion into VSV-G of an scFv peptide

Virus Exp 1 Exp 2 Exp 3 Exp 4 Exp 5 Exp 6 Exp 7
αMHC/VSV-G 7200 3020 3620 2880 1400 1720 1800
αHEL/VSV-G 1120 620 760 380 260 360 420
  1. Supernatant from HIV-1-producing 293T cells were passed through a 0.45 μm filter (Sarstedt). αMHC/VSV-G and αHEL/VSV-G HIV-1 samples were concentrated 100 times by centrifugation (25 kRPM at 4°C for 2 hours in a BeckmanCoulter ultracentrifuge) and were resuspended in 1% bovine serum albumine. VSV-G pseudotyped HIV-1-derived viral particles were directly used without prior concentration to infect target cells. When required, the virus was stored at -80°C. 50% confluent target cells, either human 293T (depicted) and HeLa cells, mouse Mus Dunni cells or monkey Cos-7 cells (not shown), were cultured with dilutions of virus for 16 hours in the presence of 5 μg/ml polybrene. 48 hours later, green fluorescent colonies were counted. Titres (colony forming units (cfu)/ml) on infected 293T cells of αMHC/VSV-G and αHEL/VSV-G particles from 7 independent experiments are shown. VSV-G pseudotyped HIV-1 was used as control for successful virus production and infection, and generally gave titers of 107-106 cfu/ml (data not shown).