TIM-family Proteins Promote Infection of Multiple Enveloped Viruses through Virion-associated Phosphatidylserine

Jemielity, Stephanie; Wang, Jinyize J.; Chan, Ying Kai; Ahmed, Asim A.; Li, Wenhui; Monahan, Sheena; Bu, Xia; Farzan, Michael; Freeman, Gordon J.; Umetsu, Dale T.; DeKruyff, Rosemarie H.; Choe, Hyeryun

VP40-GFP VLPs were produced in 293T cells by cotransfecting a pCAGGS-based plasmid encoding a previously described GFP-fusion version of the EBOV VP40 matrix protein [47] , a gift from Christopher F. Basler at Mount Sinai School of Medicine, with a plasmid encoding mucin-domain-deleted EBOV GP at 3:1 ratio. To make VP40-GFP VLPs bearing no entry protein, the VP40-GFP plasmid was similarly cotransfected with an empty plasmid. VLP-containing culture supernatants were harvested at 36 h post-transfection and cell debris removed by two consecutive centrifugations at 900 g. The presence of fluorescent filovirus-like particles in the supernatants was confirmed by fluorescence microscopy.