Journal: The Journal of Experimental Medicine
Article Title: Extracellular matrix protein Matrilin-4 regulates stress-induced HSC proliferation via CXCR4
doi: 10.1084/jem.20151713
Figure Lengend Snippet: Matn4 is highly expressed in HSCs but dispensable for steady-state hematopoiesis. (A) Matn2, 3, and 4 mRNA expression in LT-HSCs (Lin − Sca-1 + c-Kit + CD150 + CD48 − CD34 − ), short term HSCs (ST-HSCs; Lin − Sca-1 + c-Kit + CD150 + CD48 − CD34 + ), and multipotent progenitors (MPPs; Lin − Sca-1 + c-Kit + CD150 + CD48 + ) of WT mice. (B) Representative LSK FACS plots showing percentage of LSK in WT, Matn4 −/− , and Matn1–4 −/− mice. (C) Absolute numbers of LSK and LSKCD150 + CD48 − of WT, Matn4 −/− , and Matn1–4 −/− mice determined by FACS and normalized to the total BM cell counts per femur. (D) Frequency of B cells (B220 + ), megakaryocytes (CD41 + ), granulocytes (CD11b + Gr-1 + ), T cells (CD4 + CD8 + ), and erythrocytes (Terr119 + ) in BM of WT, Matn4 −/− , and Matn1–4 −/− mice determined by FACS. (E and F) Representative FACS plots (E) and quantification (F) of cell cycle distribution (Ki67/Hoechst) of WT, Matn4 −/− , and Matn1–4 −/− HSCs (LSKCD150 + CD48 − ). (G) Matn1, 2, and 3 mRNA expression levels of HSCs (LSKCD150 + CD48 − ) isolated from WT and Matn4 −/− mice isolated 4 wk after transplantation, derived from microarray analysis. n = 3 mice/group. Unpaired Student’s t test analysis was performed on three independent experiments. *, P < 0.05; **, P < 0.01. Data are mean ± SEM.
Article Snippet: BM cells were stained for HSCs and progenitors using antibodies CD4 (GK1.5), CD8 (53-6.7), CD11b (M1/70), Gr-1 (RB6-8C5), B220 (RA3-6B2), and TER119 (TER119); CD16/32 (93), CD41 (MW Reg 30), and CD105 (MJ7/18); and cKit (2B8), Sca-1 (D7), CD150 (TC15-12F12.1), CD48 (HM48-1), and CD34 (RAM34; eBioscience).
Techniques: Expressing, Isolation, Transplantation Assay, Derivative Assay, Microarray
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