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. 2020 Mar 6;10:48. doi: 10.3389/fcimb.2020.00048

Figure 2.

Figure 2

Site-specific mutations, disrupting the N-linked glycosylations in GP1, result in the loss of steric shielding by the EBOV-GP. Flow cytometry analysis shows steric shielding effect over HLA-I (upper panels) and MICA (lower panels) ligands by wild-type and mutated YFP-fused GPs. HEK293T cells were transfected with wild-type GP or with site-specific mutants of GPs with altered N-glycosylation sites (MLD, GCD, or both domains) fused to YFP. Thirty hours post transfection, cells (without fixation or permeabilization) were co-stained with mAbs specific for HLA-I or MICA. Live cells (7AAD negative) are shown in the contour plots. Q3/Q2 values in the panels are used for evaluating the shielding of MHC-I or MICA by the EBOV GP. End right panels show summary of Q3/Q2 values depicted in the bar chart. Results shown are from one representative analysis of three independent flow cytometry analyses. Graphs representing mean ± SEM. Two-way ANOVA Sidak's multiple comparisons (***P < 0.005, ***P < 0.0005, ****P < 0.0001).