sialic acids (sas) of cell surface glycoproteins and glycolipids  (SAS institute)

Journal: Pathogens

Article Title: Relationship Between Cell Surface Viral Glycoprotein Expression and Resistance of Parainfluenza Virus Persistently Infected Cells to Complement-Mediated Lysis

doi: 10.3390/pathogens14080815

Figure Lengend Snippet: Cell surface sialic acid levels on PI cells are similar to naïve cells. ( A , B ) Naïve and PI H1975 cells were plated in a 24-well plate. Naïve cells were mock infected as a negative control or acutely infected with PIV5 at an MOI of 10 PFU/cell. The cells were then stained with an anti-PIV5 HN monoclonal antibody and processed via flow cytometry. ( C ) Mock-infected, AI, and PI cells were treated with 1 U/mL Clostridium perfringens neuraminidase (NA) immediately following infection. At 24 hpi, cells were stained with the lectin MAL II followed by conjugated streptavidin, then analyzed via flow cytometry. MFI = mean fluorescence intensity. ( D ) Mock-infected, AI and PI cells were treated with NA immediately following infection. At 24 hpi, cells were left untreated, treated with 10% HI NHS, or treated with 10% NHS. Red fluorescent images were captured and quantified using the IncuCyte as described in . *** and **** show p -values of <0.001 and <0.0001.

Article Snippet: Cell surface sialic acid was monitored by flow cytometry using MALL II Maackia Amurensis Lectin II that was biotinylated (1:50, B-1265-1, Vector Laboratories, Newark, CA, USA) and then with treatment with Pacific Blue conjugated Streptavidin (1:1000, S11222 , Thermo Fisher Scientific, Waltham, MA, USA).

Techniques: Infection, Negative Control, Staining, Flow Cytometry, Fluorescence