Journal: Journal of Virology

Article Title: Mechanical Barriers Restrict Invasion of Herpes Simplex Virus 1 into Human Oral Mucosa

doi: 10.1128/JVI.01295-17

Figure Lengend Snippet: Nectin-1 and HVEM are expressed in human oral epithelia and primary keratinocytes. (a) RNAs were isolated from epithelial sheets prepared from vestibular mucosa or from primary oral keratinocytes. RT-PCR demonstrates expression of nectin-1 and HVEM; as a control, GAPDH transcripts are shown. (b) Cells were dissociated from vestibular epithelial sheets by TrypLE Select treatment and stained for nectin-1. To investigate surface expression of HVEM, cells were dissociated from epithelial sheets by CDS treatment. Flow cytometric analyses indicate 92% and 40% of cells were positive for nectin-1 and HVEM, respectively. APC-A, allophycocyanin-conjugated antibody; FITC-A, fluorescein isothiocyanate-conjugated antibody. (c) Results from samples from 4 (nectin-1) or 3 (HVEM) different patients are shown as means ± standard deviation (SD), demonstrating that surface expression of HVEM is present on fewer cells than nectin-1.

Article Snippet: Cell suspensions prepared by TrypLE Select (Life Technologies) were incubated in PBS–5% FCS on ice for 45 min with mouse anti-nectin-1 (CK41; 1:100) ( 40 ), and nectin-1 was visualized with anti-mouse IgG-Cy5 (1:100 [Jackson ImmunoResearch Laboratories, Inc.]).

Techniques: Isolation, Reverse Transcription Polymerase Chain Reaction, Expressing, Control, Staining, Standard Deviation