Journal: bioRxiv

Article Title: The anti-HIV Drug Nelfinavir Mesylate (Viracept) is a Potent Inhibitor of Cell Fusion Caused by the SARS-CoV-2 Spike (S) Glycoprotein Warranting further Evaluation as an Antiviral against COVID-19 infections

doi: 10.1101/2020.04.24.060376

Figure Lengend Snippet: Vero cells were transfected with plasmid vectors expressing either the S-o or S-n glycoproteins tagged with the 3xFLAG and N-myc epitopes at their amino termini, respectively. S-n and S-o expression was detected with mAbs against the epitope tags at 48 hours post transfection and compared to vehicle containing equivalent amount of lipofectamine. Methanol fixed cells were incubated with mouse anti-N-myc (S-n) (1:500 or 1:50) or mouse anti-FLAG (S-o) (1:2500 or 1:200) antibody and stained with either (A) HRP staining or (B) Alexa fluore 647 conjugated goat anti-mouse secondary antibody (1:1000). Cellular tubulin was stained with rabbit anti-alpha tubulin (abcam; 1:200) and anti-rabbit secondary antibody conjugated with alexa fluore 488. DAPI was used to stain nuclei of cells. Phase contrast images were taken at X10 magnification, whereas the fluorescent images were taken at 40X magnification.

Article Snippet: Cells were stained for FLAG (S-o, anti-FLAG-1:2500) or N-myc (S-n, anti-myc-1:500) with HRP (Vector Nova Red kit) for phase contrast microscopy.

Techniques: Transfection, Plasmid Preparation, Expressing, Incubation, Staining

Journal: bioRxiv

Article Title: The anti-HIV Drug Nelfinavir Mesylate (Viracept) is a Potent Inhibitor of Cell Fusion Caused by the SARS-CoV-2 Spike (S) Glycoprotein Warranting further Evaluation as an Antiviral against COVID-19 infections

doi: 10.1101/2020.04.24.060376

Figure Lengend Snippet: (A) Vero cells were transfected with plasmids expressing either the S-o or S-n glycoproteins tagged with the 3xFLAG and N-myc epitopes at their amino termini, respectively. S-n and S-o expression was detected with mAbs against the epitope tags at 48 hours post transfection and compared to vehicle containing equivalent amount of DMSO. (B) S-n and S-o glycoproteins were expressed as in A. Nelfinavir was added at the time of transfection at the concentrations indicated. Methanol fixed cells were incubated with mouse anti-N-myc (S-n) or mouse anti-FLAG (S-o) antibody and stained with HRP staining followed by goat anti-mouse secondary antibody incubation. Images were taken at X10 magnification.

Article Snippet: Cells were stained for FLAG (S-o, anti-FLAG-1:2500) or N-myc (S-n, anti-myc-1:500) with HRP (Vector Nova Red kit) for phase contrast microscopy.

Techniques: Transfection, Expressing, Incubation, Staining