Figure 2. Characterization of the hematopoietic compartment in the BM of STZ-treated animals.

(A) Number of LSK per femur defined by flow cytometry in STZ-treated or control mice. Data are mean ± s.e.m., n=24, *** p<0.001. (B) CFU-C numbers of BM cells obtained from STZ-treated and control mice. Data are mean ± s.e.m., n=6, * p<0.05. (C) Number of LT-HSC Lin⁻Sca⁺cKit⁺CD150⁺CD48⁻ and (D) Lin⁻Sca⁺cKit⁺Flk2⁻CD34⁻ per femur in STZ-treated or control mice as assessed by flow cytometry. Data are mean ± s.e.m., n=8, **p<0.01 and * p<0.05 respectively. (E–F) Histogram plots showing the fraction of LSK (E) and LT-HSC (F) in different phases of the cell cycle in STZ-treated versus control mice. Coloumns are mean ± s.e.m., n=4, * p<0.05. (G) Schematic representation of the transplantation experiment to assess the number/functionality of BM cells from STZ-induced diabetic mice. (H) Donor-derived CD45.2⁺ cell engraftment 4, 8, 12 and 16 weeks after transplantation in the peripheral blood of secondary SJL recipient mice transplanted with bone marrow isolated from STZ-treated or control mice and mixed with equal amount of CD45.1 competitor cells. Columns represent mean ± s.e.m., n=10, *** p<0.001. (I) CFU-C (relative to input) mean number ± s.e.m of the migrated purified LSK from STZ-treated or control mice toward CXCL12 (300ng/ml), n=6, *** p<0.001. (J) FACS-sorted LSK from control or STZ-treated mice were plated onto 24-well plates previously coated with 10μg/ml fibronectin. After 4 hours the adhesive LSK content was plated into methylcellulose. Data are mean CFU-C number relative to input ± s.e.m., n=6, *** p<0.001. (K) Representative FACS plots and bars indicating delta mean fluorescence intensity (DMFI) over controls showing the expression levels of CXCR4, α₄ integin, α₅ integin, and L-selectin in LSK isolated from STZ-treated and control mice, n=9, * p<0.05.