Skip to main content
. 2023 Oct 27;26(11):108299. doi: 10.1016/j.isci.2023.108299

Figure 2.

Figure 2

Impact of BA.2.12.1 and BA.4/5 mutations on Spike-mediated infection

(A) Infection kinetics of Caco2, A549-ACE2, and A549-ACE2-TMPRSS2 cells infected by VSVpp carrying the indicated mutant S proteins. Infected GFP+ cells were automatically quantified over a period of 22 h. Exemplary kinetic representing one out of four independent experiments (from Figure 2B), where each dot indicates the mean of technical duplicates.

(B) Automatic quantification of infection events by counting GFP positive cells of Caco2, A549-ACE2, and A549-ACE2-TMPRSS2 cells infected by VSVpp carrying SARS-CoV-2 Spike protein of Hu-1 (gray), BA.1 (light green), BA.2 (yellow), BA.2.12.1 (orange), or BA.4/5 (dark red). Bars represent the mean of four independent experiments (±SEM). Statistical significance was tested by one-way ANOVA. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

(C) Automatic quantification of infection events of Caco2 (upper panel), A549-ACE2 (middle panel), and A549-ACE2-TMPRSS2 (lower panel) cells transduced with VSVΔG-GFP pseudotyped with SARS-CoV-2 S of Hu-1 (gray), BA.1 (light green), BA.2 (yellow), BA.2.12.1 (orange), or BA.4/5 (dark red) or indicated mutant S proteins. Graphs in the center show Hu-based S mutants, infection events are normalized on the Hu-1 S (set at 1) and p values indicate difference to the Hu-1 S. Graphs on the right show BA.2-based S mutants and infection events are normalized on the BA.2 S (set at 1) and p values indicate difference to the BA.2 S. Bars represent the mean of four independent experiments (±SEM). Statistical significance was tested by one-way ANOVA. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001.

(D) Correlation of the pseudotype infection events of the wild-type S or indicated mutant S proteins in A549-ACE2-TMPRSS2/A549-ACE2 (upper panel), A549-ACE2/Caco2 (middle panel), and A549-ACE2-TMPRSS2/Caco2 (lower panel) cells. Each symbol represents the average value derived from four infection experiments (shown in Figure 2C). Numbers indicate the combined mutations of BA.4/5 S proteins whose infection events do not correlate between A549 and Caco2 cells. Coefficient of correlation (r), coefficient of determination (R2-values) and two tailed p values are provided (see also Figure S2).