Fig. 5

Flow cytometry analysis of mouse bone marrow mesenchymal stem cells. Freshly isolated mouse total bone marrow (BM) cells were analysed by flow cytometry for identifying (A-E) mesenchymal stem cells (MSC) (gating for CD45, CD11b, CD29, Sca-1 and CD73), and next for detecting senescence (gating for CD36 and β-galactosidase Senescence Green) (F&H), mobilization potential (gating for DPP4 (CD26)) (G) and mitochondrial activity in living cells (gating for MitoTracker Deep red) (I). ​The gating strategies consisted of selecting live singlet populations (Ai-Aiii), sorting for CD45^negCD11b^negCD29^pos MSCs (Aiv-Av), followed by examining the ratio of cell fractions positive for Sca-1, CD73, DPP4, and CD36 (Avi-Avii), and finally detecting the median fluorescence intensity (MFI) of β-galactosidase Senescence Green and MitoTracker Deep red in BM-MSCs. Representative histograms (Bi-Bv) showing markers expression in BM-MSCs (red) and their FMO controls (blue). Fluorescence intensity of β-galactosidase Senescence Green data is displayed on Bi-exponential plots. This is a hybrid scale where it is logarithmic for both positive and negative values until the 1st decade (-10 to + 10) which is displayed as linear. Bar graphs show individual values and means ± SEM. n = 4 to 6 animals per group