Journal: Cell stem cell

Article Title: Regulation of hematopoietic stem and progenitor cell mobilization by cholesterol efflux pathways

doi: 10.1016/j.stem.2012.04.024

Figure Lengend Snippet: Plasma IL-17 levels in WT and Abca1−/−Abcg1−/− recipient mice transplanted with either WT or Abca1−/−Abcg1−/− BM 8 weeks post-reconstitution (A). Plasma G-CSF levels in WT recipients transplanted with WT and Abca1−/−Abcg1−/− BM 14 weeks post-reconstitution and i.p injected with IgG control or 250µg IL-17 neutralizing antibody for 16h (B). Colony-forming unit numbers from the blood of these mice (C). Plasma IL-17 levels in WT and Abca1−/−Abcg1−/− mice treated with broad-spectrum antibiotics for 2 weeks to deplete commensal bacteria (D). IL-23 protein content in the spleen of WT and Abca1−/−Abcg1−/− recipient mice transplanted with either WT or Abca1−/−Abcg1−/− BM 8 weeks post-reconstitution (E). Plasma IL-17 levels in WT recipients transplanted with WT and Abca1−/−Abcg1−/− BM 14 weeks post-reconstitution and i.p injected with IgG control or 200µg IL-23R neutralizing antibody for 16h (F). Results are ± SEM of 5 to 6 animals per group. *P<0.05 vs. WT recipients transplanted with WT BM. #P<0.05 vs. IgG control. §P<0.05 vs. Abca1−/−Abcg1−/− recipients transplanted with Abca1−/−Abcg1−/− BM.

Article Snippet: For neutralizing antibody experiments, WT and Abca1 −/− Abcg1 −/− BM transplanted mice were i.p injected 16h before analysis with the following antibodies: anti-IL-3Rβ AF549, anti-Cxcl2 MAB452, anti-IL-23R MAB1686, anti-IL-17 MAB421 and anti-G-CSF MAB414 (all from from R&Dsystems).

Techniques: Injection

Journal: Cell stem cell

Article Title: Regulation of hematopoietic stem and progenitor cell mobilization by cholesterol efflux pathways

doi: 10.1016/j.stem.2012.04.024

Figure Lengend Snippet: Splenic IL-23 protein content (A), Colony-forming unit numbers (B), plasma G-CSF levels (C), and plasma IL-17 levels (D) in 28 weeks old Abca1fl/flAbcg1fl/fl (controls), LysM-Cre Abca1fl/flAbcg1fl/fl, and CD11c-Cre Abca1fl/flAbcg1fl/fl mice. Results are ± SEM of 5 to 6 animals per group. *P<0.05 vs. controls. Modulation of IL-23 mRNA expression levels in Abca1fl/flAbcg1fl/fl and LysM-Cre Abca1flfl-Abcg1fl/fl BM-derived macrophages (E) and CD11c-Cre Abca1fl/flAbcg1fl/fl BM-derived dendritic cells (F) after manipulation of plasma membrane cholesterol. Cells were incubated with 5mmol/L cyclodextrin (CD) for 30minutes before treatment with 50ng/mL lipopolysaccharide (LPS, TLR4 ligand) or 2,5µg/mL PolyI:C (TLR3 ligand) for 3 hours. IL-23 transcript levels were normalized to m36B4 mRNA amount. RNA levels were expressed as arbitrary units (a.u). Values are mean ± SEM of an experiment performed in quadruplicate. *P<0.05 vs. respective controls. §P<0.05 vs. conditions without cyclodextrin treatment.

Article Snippet: For neutralizing antibody experiments, WT and Abca1 −/− Abcg1 −/− BM transplanted mice were i.p injected 16h before analysis with the following antibodies: anti-IL-3Rβ AF549, anti-Cxcl2 MAB452, anti-IL-23R MAB1686, anti-IL-17 MAB421 and anti-G-CSF MAB414 (all from from R&Dsystems).

Techniques: Expressing, Derivative Assay, Incubation

Journal: Cell stem cell

Article Title: Regulation of hematopoietic stem and progenitor cell mobilization by cholesterol efflux pathways

doi: 10.1016/j.stem.2012.04.024

Figure Lengend Snippet: Quantification of BM macrophage subsets by flow cytometry in WT and Abca1−/−Abcg1−/− recipient mice transplanted with either WT or Abca1−/−Abcg1−/− BM 8 weeks post-reconstitution (A and B) and in WT recipients transplanted with WT and Abca1−/−Abcg1−/− BM 14 weeks post-reconstitution and i.p injected with IgG control or 250µg IL-17 neutralizing antibody for 16h (C and D). Results are ± SEM of 5 to 6 animals per group. *P<0.05 vs. WT. §P<0.05 vs. Abca1−/−Abcg1−/− recipients transplanted with Abca1−/−Abcg1−/− BM. Quantification of hematopoietic progenitors (E), monocytes and neutrophils (F) in WT BM cultures grown for 48h in liquid culture in presence of 10% of the indicated serum and 50ng/mL G-CSF neutralizing antibody (+) or control non-specific IgG (−). Results are ± SEM of 3 independent experiments. #P<0.05 vs. IgG control. †P<0.05 vs. 10% WT serum.

Article Snippet: For neutralizing antibody experiments, WT and Abca1 −/− Abcg1 −/− BM transplanted mice were i.p injected 16h before analysis with the following antibodies: anti-IL-3Rβ AF549, anti-Cxcl2 MAB452, anti-IL-23R MAB1686, anti-IL-17 MAB421 and anti-G-CSF MAB414 (all from from R&Dsystems).

Techniques: Flow Cytometry, Injection