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. 2017 Sep 18;114(40):E8508–E8517. doi: 10.1073/pnas.1712592114

Fig. 3.

Fig. 3.

(A) MERS-CoV S1-Fc hemagglutination phenotype is dependent on multivalent presentation on nanoparticles and on Sia-containing receptors on the erythrocyte surface. Human erythrocytes were mock-treated (PBS) or NA-treated, and incubated with a twofold serial dilution of MERS-CoV S1-Fc, pA-LS, or a combination. Hemagglutination was scored after 2 h of incubation at 4 °C. Wells positive for hemagglutination are encircled. Experiments were performed at least two times. (B) Defining the optimal pA-LS/S1-Fc ratio for hemagglutination. A fixed amount of MERS-CoV S1-Fc (2.5 μg) was preincubated with varying amounts of pA-LS (4, 2, 1, 0.5, 0.25, or 0.125 μg) serially diluted twofold and then incubated with washed human erythrocytes. Hemagglutination was scored after 2 h of incubation at 4 °C, and an optimal pA-LS/S1-Fc ratio was determined. Conditions lacking either pA-LS or MERS-CoV S1-Fc protein were used as a negative control. Wells positive for hemagglutination are encircled. Experiments were performed at least two times.