Journal: PLOS ONE

Article Title: Human BST2 inhibits rabies virus release independently of cysteine-linked dimerization and asparagine-linked glycosylation

doi: 10.1371/journal.pone.0292833

Figure Lengend Snippet: (A) HEK293T cells were transfected with plasmids expressing flag-hBST2. Cell lysates were prepared and incubated in the presence or absence of PNGase F and subjected to western blot analysis of hBST2. HEK293T cells were transfected with (B) 1 μg of flag-hBST2 plasmid and an increasing amount (0.5, 1 and 2 μg) of plasmids encoding RABV G-myc, M-myc, N-myc and P-myc or (C) 1 μg of plasmids encoding individual myc-tagged RABV proteins with an increasing amount of flaghBST2 plasmid (0.5, 1 and 2 μg). At 48 hpt, cell lysates were collected and analyzed by western blotting for RABV protein and hBST2 expression using rabbit α-myc and α-human BST2 antibodies, respectively. Arrow heads indicate glycosylated monomeric hBST2 of approximately 30 kDa. Arrows indicate a low molecular weight, glycosylated dimeric hBST2 of approximately 55 kDa. Asterisks indicate high molecular weight glycosylated dimeric hBST2 of approximately 65 kDa.

Article Snippet: HEK293T cells seeded in 10-cm 2 dishes were transfected with 6 μg of plasmid expressing RABV Gmyc and flaghBST2 complexed with Fugene ® HD transfection reagent (Promega).

Techniques: Transfection, Expressing, Incubation, Western Blot, Plasmid Preparation, Molecular Weight, High Molecular Weight

Journal: PLOS ONE

Article Title: Human BST2 inhibits rabies virus release independently of cysteine-linked dimerization and asparagine-linked glycosylation

doi: 10.1371/journal.pone.0292833

Figure Lengend Snippet: HEK293T cells were transfected with plasmids expressing RABV Gmyc and flaghBST2. At 24 hpt, cells were fixed and probed with mouse α-myc and rabbit α-human BST2 primary antibodies to detect RABV G and hBST2, respectively followed by Alexa Fluor ® 488-conjugated goat α-rabbit IgG H&L and Alexa Fluor ® 647-conjugated goat α-mouse IgG H&L secondary antibodies. Localization of the proteins were visualized by confocal microscopy.

Article Snippet: HEK293T cells seeded in 10-cm 2 dishes were transfected with 6 μg of plasmid expressing RABV Gmyc and flaghBST2 complexed with Fugene ® HD transfection reagent (Promega).

Techniques: Transfection, Expressing, Confocal Microscopy