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sc-12-1058_sm_SupplFigure1.pdf228KFigure S1. Sca-1+/CD45−/CD31- and CD45+ cell proportions in different fat depots. (A-C) Percentage or cell number per fat depot (D) of Sca-1+/CD45−/CD31- and CD45+ cells in the SVF from ScAT (A), PGAT (B) and MesAT (C). Results are expressed as mean ± SEM of n≥10 separate experiments. * p< 0.01 and ** p<0.001 when compared to ScAT. (E) Representative image of lipid droplet staining with BODIPY Dye in CD45+ cells after 10 days in adipogenic culture conditions.
sc-12-1058_sm_SupplFigure2.tif1491KFigure S2. AMD3100 treatment does not modify murine ASC viability and phenotype. (A) Percentage of dead cells in the SVF from SCAT, PGAT and MesAT treated or not with AMD3100 (10−6M) in vitro for 180 min. (B) Percentage of Sca-1+/CD45−/CD31- cells in the SVF from SCAT, PGAT and MesAT treated or not with AMD3100 (10−6M) in vitro for 180 min.
sc-12-1058_sm_SupplFigure3.tif1664KFigure S3. In vivo, the administration of CXCR4 antagonist AMD3100 does not increase the number of Sca-1+/CD45−/CD31- cells in blood. (A-C) Representative dot blots showing flow cytometry cellular staining profiles of blood samples from 10 week-old C57BL/6 mice. (A) Forward-scattered light (FSC) is proportional to cell-surface area or size and side-scattered light (SSC) is proportional to cell granularity or internal complexity. (B, C) Representative dot blots showing flow cytometry staining profiles into the CD45- cell population with Sca-1 and CD31 antibodies in mice treated (C) or not (B) avec AMD3100.
sc-12-1058_sm_SupplFigure4.pdf291KFigure S4. Lymph node structure and injected GFP+-ASCs localization. (A) Representative image of PLNs from mice injected in the footpad with GFP+ ASCs (in green) 12 hours before sacrifice and stained Hoescht 33342 (in blue). (B-D) Representative images of PLNs slices stained with (B) anti-B220 (red), (C) anti-CD3 (red) or anti-ER-TR7 (red) and Hoescht 33342 (blue). Representative microscopy results of at least 3 independent experiments.
sc-12-1058_sm_SupplFigure5.pdf335KFigure S5. T cell activation markers increase in OVA-IFA immunized draining lymph node while no changes are detected in BM cell content. Mice were immunized subcutaneously with 50μg of ovalbumin (OVA) in IFA in the right footpad. Left footpad was injected with saline. Nine days later, draining lymph node, popliteal (right) and inguinal (left) were separately harvested. (A) Lymph node cells were labelled with antibody cocktail. CD44 and CD62L expression gated on TCRb+CD4+ cells and CD44 and CD62L expression gated on TCRb+CD8+ cells. (B,C) The femurs from control or vaccinated hindlimbs were harvested and BMs were recovered by flushing. Total cell number was determined (B) (see methods) and the Sca- 1+/CD45−/CD31- cell content was determined by flow cytometry and compared to the one of control femurs, 9 days after the immunization procedure. Results are expressed as mean ± SEM of n=6 separate experiments.
sc-12-1058_sm_SupplVideo1.avi52228KSupplementary Video 1
sc-12-1058_sm_SupplVideo2.avi2355KSupplementary Video 2

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