Article Title: Signal-transducing adapter protein-1 is required for maintenance of leukemic stem cells in CML
Figure Lengend Snippet: STAP-1 does not affect normal murine hematopoiesis. a The indicated subsets of hematopoietic cells derived from WT C57BL/6 mice were analyzed for STAP-1 expression by real-time RT-PCR ( n = 3). STAP-1 transcript levels were normalized to myeloid subset. b Blood counts (WT, n = 5; STAP-1 KO, n = 7) and biochemical parameters (WT, n = 4; STAP-1, KO n = 4) were evaluated using peripheral blood. c The frequency of LSK and LT-HSC (CD48 − CD150 + LSK) cells from WT and STAP-1 KO mice was analyzed. Representative flow cytometry plots for identification of LT-HSC were shown (left panels). Bar graph represents mean proportion of LSK cells or LT-HSCs (WT, n = 9; STAP-1 KO, n = 8). d LSK cells from WT or STAP-1 KO BM ( n = 6) were cultured in methylcellulose medium for 7 days, colonies were counted, and cells were serially replated. e WT or STAP-1 KO LSK cells (5000 cells, Ly5.2) were transplanted into lethally irradiated recipients (Ly5.1). At 4 months after transplantation, 2 × 10 6 whole BM cells from the primary recipient mice were transplanted into the secondary recipient mice (Ly5.1). The left line graphs represent the mean percentages of donor chimerism in recipient’s PB at the indicated time points after transplantation (WT, n = 9; STAP-1 KO, n = 7 for the first BMT, and n = 11 per group for the second BMT). Upper bar graphs represent chimerism of donor-derived cells in BMMNC cells and LSK cells from primary BMT recipients of WT or STAP-1 KO LSK cells 4 months after primary BMT (WT, n = 9; STAP-1 KO, n = 7). Lower bar graph represents differentiation status (B220 + B cells, CD3 + T cells, or Mac-1/Gr-1 + myeloid cells) in BM cells from primary BMT recipients of WT or STAP-1 KO LSK cells 4 months after primary BMT (WT, n = 9; STAP-1 KO, n = 7). Data shown represent the mean ± SD. * p
Article Snippet: Antibodies against STAP-1 (rabbit polyclonal) were purchased from Sigma-Aldrich.
Techniques: Derivative Assay, Mouse Assay, Expressing, Quantitative RT-PCR, Flow Cytometry, Cell Culture, Irradiation, Transplantation Assay