The native and engineered RBCs were plated in 96-well plates at 1 × 105 cells per well in 100 μl of 10 mM PBS. Then, 10 μl of anti-D, anti-A, and/or anti-B blood grouping reagent (monoclonal antibodies, SHPBC, China) was added to each well, and the cells were incubated for 1 hour at room temperature. Incubated cells were stirred using pipette tips and then observed using optical microscopy, and the images were captured on an Axio Observer A1 microscope (Zeiss, Germany).

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