Journal: Nature medicine
Article Title: Bone marrow-derived immature myeloid cells are a main source of circulating suPAR contributing to proteinuric kidney disease.
doi: 10.1038/nm.4242
Figure Lengend Snippet: Figure 3 BM immature myeloid cells have an ability to transfer disease. (a) Examination of proteinuria in recipient NSG mice before (0) and 6, 12, and 24 h after adoptive transfer of BM cells obtained from PBS- or LPS-injected NSG mice. The results are from two independent experiments (n = 4 for PBS-BMCs; n = 7 for LPS-BMCs). Data are shown as mean ± s.e.m.; unpaired two-tailed Student t-test, *P < 0.05. (b) Examination of proteinuria in recipient NSG mice 12 h following adoptive transfer of BM cells obtained from LPS-challenged C57BL/6 WT and Pod-Rac1 mice. n = 5 per group. Data are shown as mean ± s.e.m. One-way ANOVA, Tukey’s multiple comparison test, **P < 0.01, NS, not significant. (c) Representative dot plots (chosen from a total of three generated) of triple-color stained (uPAR/Sca-1/Gr-1) total BM cells (red) isolated from PBS- or LPS-injected C57BL/6 mice (n = 3 per group). uPAR+ cells (blue) were gated and shown in these dot plots. (d) Quantitation for uPAR-expressing immature myeloid cells (uPAR+Sca-1loGr-1lo) cells shown in c (n = 3 per group). Data are shown as mean ± s.e.m.; unpaired two-tailed Student t-test, ***P < 0.001. (e–g) In vitro culture of total BM cells isolated from C57BL/6 WT mice with PBS or various concentrations of LPS (0.1, 1, and 10 µg/ml). (e) Experimental scheme. (f) In vitro induction of uPAR+Sca-1loGr-1lo cells determined by triple color-flow cytometric analysis. (g) suPAR secretion into culture medium (CM) measured by suPAR ELISA. Data are shown as mean ± s.e.m. One-way ANOVA, Tukey’s multiple comparison test, *P < 0.05, **P < 0.01, ***P < 0.001. (h) Schematic diagram outlining the experimental design of the BM cell transfer used in i–k. (i–k) Examination of proteinuria (i), serum (j) and urinary suPAR levels (k) in recipient NSG mice before (0) and 6, 12, and 24 h after adoptive transfer of either whole BM cells or Sca-1+ cell-depleted (Sca-1∆) BM cells of LPS-challenged C57BL/6 WT mice. The results are from three independent experiments (n = 9 for whole BMCs; n = 8 for Sca-1∆BMCs). Data are shown as mean ± s.e.m.; unpaired two-tailed Student t-test, *P < 0.05.
Article Snippet: Mouse suPAR levels from plasma (and/or serum) and urine were evaluated by enzyme-linked immunosorbent assay (ELISA) kit (R&D systems, DY531) following the manufacturer’s protocol.
Techniques: Adoptive Transfer Assay, Injection, Two Tailed Test, Comparison, Generated, Staining, Isolation, Quantitation Assay, Expressing, In Vitro, Enzyme-linked Immunosorbent Assay
R&D Systems is a verified supplier 
R&D Systems manufactures this product 
