Extended Data Figure 7: Overexpression of S1P₁ results in marked decreases in lymphocyte populations in the thymus and secondary lymphoid organs.

a, Representative flow cytometry plots and quantitative MFI of S1P₁ expression by Lin⁻ cells from S1pr1 OE and wild-type (WT) littermates. n = 3. b, Relative expression levels of S1pr1 mRNA in bone marrow cells of S1pr1 OE mice relative to wild-type mice, as determined by multiplex qRT–PCR. c, Representative agarose gel of Cre activation and excision of the floxed stop cassette as assessed by PCR of genomic DNA from bone marrow cells of S1pr1 OE mice, lox/stop/lox littermate, or wild-type littermate. Arrows indicate amplified DNA fragments corresponding to undeleted or deleted segments d–f, Four days after the final dose of tamoxifen, CD4⁺, CD8⁺ and CD19⁺ cells in brachial and inguinal lymph nodes (d) and spleen (e), and thymic CD4⁺CD8⁺ double-positive (DP), CD4⁺ and CD8⁺ single-positive (CD4 or CD8 SP), total double-negative (DN), and double-negative subpopulations DN2, DN3 and DN4 (f) were quantified by flow cytometry in S1pr1 OE mice and wild-type littermates. Bars represent means ± s.d.; S1pr1 OE, n = 6; wild type, n = 7 and data are compiled from two experiments.