Figure 5.

Validation of mutations in chemokine receptors that increase BiFC.A and B, mutations of CXCR4 predicted from the deep mutational scan to increase BiFC with WT CXCR4 were validated. The mutations were introduced into CXCR4-VC and transfected into X4-KO Expi293F cell lines stably expressing either (A) CXCR4-VN or (B) mGluR3-VN. C and D, validation of CCR5 mutants enriched in the deep mutational scan for high BiFC signal. The mutations in the CCR5-VC construct were tested in cell lines stably expressing either (C) CCR5-VN or (D) mGluR3-VN. BiFC signal was measured by flow cytometry. Data are mean ± SD, n = 3 independent replicates. ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001; one-way ANOVA with Dunnett test. BiFC, bimolecular fluorescence complementation.