Figure 5

Characterisation of homotypic interactions of Ag43 variants by flow cytometry. Flow cytometry analyses were performed by mixing SRUD-Green and SRUD-Blue both expressing the same Ag43, namely Ag43 ^C1-PT (C1), Ag43^C2-PT (C2), Ag43^C3-PT (C3) or Ag43^C4-PT (C4). (A) Based on the SSC-A over FSC-A representation (pictured in the insert), the fluorescent FITC-H over PB450-H representation focused on the gate corresponding to the cell aggregates as previously defined (Supplementary Fig. S3). The green cloud corresponds to cell aggregates of SRUD-Green, the blue cloud to cell aggregates of SRUD-Blue, and the orange cloud to mixed cell aggregates of SRUD-Green and SRUD-Blue. Graphs correspond to one representative experiment. (B) The percentage of mixed cell aggregates was calculated from the events detected in orange cloud over the total number of events corresponding to aggregates. (C) The concentration of cells was calculated from the gates corresponding to single individual cells as previously defined (Supplementary Fig. S3). (D) The size of aggregates was recovered from the gate corresponding to cell aggregates as previously defined (Supplementary Fig. S3).