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. 2018 Sep 26;92(20):e00858-18. doi: 10.1128/JVI.00858-18

FIG 2.

FIG 2

T3DF/T3DCS1 attaches poorly to L929 cells. (A) Confluent monolayers of L929 cells grown in 96-well plates were adsorbed with 5 × 104 particles/cell of either T3DF or T3DF/T3DCS1 at 4°C for 1 h. Cell attachment was determined by indirect immunofluorescence of cell-associated particles using a LI-COR Odyssey scanner. Binding index was determined by calculating intensity ratios at 800 nm (green fluorescence), representing viral antigen, and 700 nm (red fluorescence), representing the cell monolayer. The binding index for each independent infection and the sample means are shown. Error bars indicate SD. *, P < 0.05 as determined by Student's t test compared to T3DF. (B) To examine the effect of 9BG5 MAb on virus infectivity, 1 × 1011 particles/ml of either T3DF or T3DF/T3DCS1 were incubated at 4°C with 0 or 500 ng/ml of 9BG5 MAb hybridoma supernatant overnight. This mixture was used to initiate infection of L929 cells in 96-well plates. (C) To examine the role of glycans on infection, L929 cells were pretreated with 0 or 10 mU/ml neuraminidase. Cells were adsorbed with 3,000 particles/cell of either T3DF or T3DF/T3DCS1 at room temperature for 1 h. (B and C) After incubation at 37°C for 18 h, the cells were subjected to indirect immunofluorescence assay using a LI-COR Odyssey scanner. Relative infectivity was determined by calculating intensity ratios at 800 nm (green fluorescence), representing viral antigen, and 700 nm (red fluorescence), representing the cell monolayer. For each virus strain, the infectivity index for untreated samples was set to 100%. Percent infectivity for each independent infection and the sample mean are shown. Error bars indicate SD. *, P < 0.05 as determined by Student's t test compared to T3DF. **, P < 0.05 compared to similarly treated T3DF-infected cells.