With both S1-shedding and large-scale virus lysis ruled out, we turned to examination of the site(s) on the S protein potentially important for the inhibitory function of lectins. As mentioned earlier, SARS-CoV-2 Spike has 22 highly conserved N-linked glycans in each of the three protomers , distributed between the S1 and S2 subunits (Figure ). To determine if the mechanism of action for irreversible inactivation by CV-N is due to a specific subset of glycans on the SARS-CoV-2 S protein, we tested single glycan attachment-site mutants of SARS-CoV-2 for continued susceptibility to lectin inhibition. After pseudovirus production, the infectivity of the glycan attachment site mutants and wild type (WT) SARS-CoV-2 pseudoviruses were measured.