Testing the importance of residues lining the extracellular facing half of TM2-TM11 and TM5-TM8 in LPC transport. (A) The model of the outward-open gMfsd2a structure (PDB: 798N). TM2 is in teal, TM11 in blue, TM5 in red, and TM8 in orange. Gray sticks represent residues that were mutated. Residues correspond to human Mfsd2a numbering. (B) Quantification of transport of indicated mutants using a single-cell Flow cytometry assay. The location of residues lining TM2-TM11 and TM5-TM8 are indicated above the graph. Data are represented as the mean ± SD, with each dot representing a single analyzed cell with approximately 1,000 cells analyzed per construct. The dotted line represents the average transport level of WT. See SI Appendix, Fig. S4 for primary flow data.