Extended Data Figure 1. Lymphoma-associated mutations result in loss of expression and function of S1PR2.

(a-c) Surface expression of FLAG (a), quantitative PCR of human S1PR2 (b) or Thy1.1 reporter expression (c) in mouse WEHI231 B lymphoma cells transduced as described in Fig. 1b. Shown in a are histograms of transduced cells (Thy1.1⁺) in blue and untransduced cells (Thy1.1⁻) in gray. 5 of 8 S1PR2 mutations showed loss of protein expression despite strong transcript and reporter expression. Loss of expression in these 5 mutants was likely a result of degradation of improperly folded proteins in the ER. (d) Representative FACS plots of transwell migration of WEHI231 cells transduced with vector, WT or R147C mutant S1PR2 to the indicated stimuli or the input sample. Numbers indicate % of cells positive for the Thy1.1 reporter. (e) WEHI231 cells stimulated as in Fig. 1d were analyzed for phosphorylation of Akt (pAkt S473) by Western blot or by intracellular FACS. Data in a and c are representative of 4 independent experiments and d and e of 3 independent experiments.