Characterization of MSC phenotype in SCD. (A-B) SCD mice were analyzed by using flow cytometry for total bone marrow cellularity (A) and Ter119^–CD45^–CD31^–CD51⁺PDGFRα⁺ MSC frequency of total bone marrow cells (B). (C) ROS mean fluorescence intensity (MFI) in MSCs was determined by DCFDA. N = 5 mice for panels A-C. Sorted MSCs from Townes SS mice in culture were differentiated into adipogenic, chondrogenic, and osteogenic lineages. (D) Representative images of Oil-red-O staining to mark lipid droplets among adipogenic differentiated cells, Alcian blue for chondrogenic differentiation, and Alizarin red for osteogenic differentiation. Scale bar, 100 µM. Images are representative from N = 6 mice. (E) Corresponding adipogenic (Fabp4, Pparγ), chondrogenic (Acan), and osteogenic (Sp7, Col1a2) specific gene expression determined by RT-PCR (N = 3-6 mice). Gene expression is relative to Gapdh. Data were analyzed with a 2-tailed, unpaired Student t test. Data are presented as mean ± standard error of the mean. *P ≤ .05, **P ≤ .01, ****P ≤ .0001.