Fig. 4

Frequency of in vivo HSC and LT-HSC hematopoietic progenitors in adult wild-type and Emilin-2 null mice. BM were isolated from wild-type and Emilin2^−/− mice of 3,6 and 10 months of age and subjected to flow cytometry with different antibodies. A Representative flow cytometry analysis of HSC and LT-HSC in BM of 10-month-old mice, based on the identification of lineage-negative (Lin⁻) cells and of Sca1, c-Kit, CD150 and CD48 markers. Live cells, identified with side scatter (SSC; granularity) and forward scatter (FSC; cell size) parameters (plot i), negative for Lin⁻marker (plot ii), were selected and analyzed for Sca1 and c-Kit signal (plot iii). Double positive Sca1⁺/c-Kit⁺ events were identified as HSC (squared areas of plot iii) and subjected to further analysis based on CD150 and CD48 signals to identify LT-HSC (Cd150⁺/CD48⁻; squared area of plot iv). B Percentages of HSC, calculated on total live cells and on Lin⁻ cells, n BM of 3-month-old (wild type, n = 4;Emilin2^−/−, n = 4), 6-month-old (wild-type, n = 10; Emilin2^−/−, n = 6) and 10-month-old (wild-type, n = 7; Emilin2^−/−, n = 9) mice. C Percentage of LT-HSC, calculated on either total live cells, Lin⁻ cells and HSC cells, in BM of 10-month-old (wild-type, n = 3; Emilin2^−/−, n = 4) mice