Sphingolipid concentration in selected tissues of inducible Sgpl1-deficient mice.

<p>Two weeks after tamoxifen induction, tissues of Sgpl1^Flox/Flox Cre^+/− mice (open bars) and of Sgpl1^Flox/Flox Cre^−/− controls (filled bars) were obtained (n = 5/group). Tissues were extracted and sphingolipids were quantified by LC/MS. <i>A, B,</i> S1P; <i>C, D</i>, Sph; <i>E,</i> C16-ceramide. <i>A, C,</i> and <i>E</i> show absolute concentrations per weight of tissue; <i>B and D</i> show fold increase in inducible KO mice.</p

Foxp3⁺ Treg are overrepresented in LN and spleen of in inducible Sgpl1-deficient mice.

<p>Two weeks after tamoxifen treatment, cells from blood, LN and spleen were stained for T cell markers and Foxp3. <i>A,</i> Mean percentage and <i>B</i>, absolute numbers of Foxp3⁺ cells among CD4⁺ T cells (n = 4/group).</p

Normal T cell development, reduced splenic cellularity, and increased LN cell number in inducible Sgpl1-deficient mice.

<p>B and T cell subpopulations in tamoxifen-treated Sgpl1^Flox/FloxCre^+/− (open bars) and Sgpl1^Flox/Flox Cre^−/− mice (closed bars) (n = 4/group), were enumerated based on total live cell counts and cell proportions as established by flow cytometry. <i>A</i>, Thymus; <i>B, C</i>, spleen; <i>D, E</i> lymph nodes. In <i>C</i> and <i>E</i>, CD8 and CD4 T cells were analysed for co-expression of CD44 and CD62L to define populations of naive and memory T cells; the insert in <i>C</i> provides a gating example for naïve/memory type T cells.</p

Protection of inducible Sgpl1-deficient mice in EAE.

<p>Tamoxifen-induced Sgpl1^Flox/Flox Cre^+/−, Sgpl1^Flox/Flox Cre^−/−, Cre^+/−, and Cre^−/− mice (n = 6–10/group) were immunized with MOG emulsified in Complete Freund’s Adjuvans. Data from one representative experiment out of three independent studies are shown. <i>A</i>, Incidence of mice with a clinical EAE score ≥1; <i>B</i>, clinical score; <i>C</i>, body weight. For histological analysis thoracic sections of spinal cord tissue from Sgpl1^Flox/Flox Cre^+/− and Sgpl1^Flox/Flox Cre^−/− mice undergoing EAE (day 24) were stained (<i>D</i>) with H&E to visualize CNS-invading cells (scale bar is 500 µm, arrows highlight areas of inflammation); <i>E</i>, for CD3⁺ T cells (scale bar is 500 µm, rectangles indicate area of magnification, where scale bar represents 100 µm); and <i>F</i>, with solochrome to assess the integrity of the myelin sheath (scale bar is 500 µm, arrows highlight areas of beginning demyeliniation).</p

Histology of inducible Sgpl1-deficient mice in EAE.

<p>For histological analysis thoracic sections of spinal cord tissue from Sgpl1^Flox/Flox Cre^+/− and Sgpl1^Flox/Flox Cre^−/− mice undergoing EAE (day 24) were stained (<i>A</i>) with H&E to visualize CNS-invading cells (scale bar is 500 µm, arrows highlight areas of inflammation); <i>B</i>, for CD3⁺ T cells (scale bar is 500 µm, rectangles indicate area of magnification, where scale bar represents 100 µm); and <i>C</i>, with solochrome to assess the integrity of the myelin sheath (scale bar is 500 µm, arrows highlight areas of beginning demyeliniation).</p