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Blood, Vol. 112, Issue 4, 1048-1055, August 15, 2008
B-lineage commitment prior to surface expression of B220 and CD19 on hematopoietic progenitor cells
Blood Mansson et al.
112: 1048
Supplemental materials for: Mansson et al
Files in this Data Supplement:
- Figure S1. Kinetics for development of B-cells from LIN−B220−CD19−FLT3+IL7R+SCA1loKITlo hCD25 subfractions in OP9 cocultures (JPG, 48.6 KB)
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Graphs display distribution of GR1+/MAC1+, CD19+ and CD19−cells of total PI−CD45+ cells (A) and relative number of cells generated (B) from LIN−B220−CD19−FLT3+IL7R+SCA1loKITlo subfractions. Coculture wells were, at indicated day stained with Fc-block (CD16/CD32, 2.4G2) followed by CD45.2 (104) APC-Alexa750, CD19 (6D5) FITC, B220 (RA3-6B2) PE-Cy7, hCD25 (BC96) APC, GR1 (RB6-8C5) PE-Cy5, MAC1 (M1/70) PE-Cy5 and PI. Cell numbers were estimated through time-limited acqusition of cells at constant flow rate.
- Figure S2. Distribution of hCD25 positiv cells in PI−CD45.2+MAC1−CD19−cells after indicated time of co-culture on OP9 stroma cells (JPG, 80.6 KB)
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For additional information, see legend to Fig. S1.
- Figure S3. Reconstitution in mice transplantated with LIN−B220−CD19−FLT3+IL7R+SCA1loKITlo hCD25 subfractions (JPG, 70.3 KB)
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Total doner reconstitution (CD45.2) and reconstitution of lineages positiv cells (A) in bonemarrow and spleen eight days after transplantation (through tail vein injections) with indicated population into irradiated (800 RAD) CD45.1 mice. Organs were harvested and single cells suspensions made prior to staining with Fc-block (CD16/CD32, 2.4G2) followed by CD45.2 (104) FITC, CD45.1 (A20) PE, MAC1 (M1/70) APC, NK1.1 (PK136) PacificBlue, CD19 (6D5) PE-Cy7, B220 (RA3-6B2) PE-Cy5 and PI. Panel (B) shows representative plots of doner (CD45.2) cells in spleen.
- Figure S4. Hierarchical clustering showing differentially expressed genes between hCD25+/−CLP subfractions (JPG, 276 KB)
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In detail, genes were selected for being expressed in either hCD25+ or hCD25−CLP (100+ expression units in either), differing >1.5 fold between hCD25+ or hCD25−CLP and differing >2 fold between LMPPsand pro-B cells.
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