Research Paper Volume 16, Issue 9 pp 7535—7552

Figure 3. Age-associated oxidative stress in ASCs. (A) Flow cytometry gating strategy used in this study to gate on CD34⁺ fraction after excluding DAPI positive (dead cells), CD45⁺ (hematopoietic lineage), and CD31⁺ (endothelial) cells for downstream analysis of ROS, mitochondrial membrane potential, senescence, and autophagy markers. (B) ROS concentration was measured in ASCs. A representative histogram of flow cytometric measurement of DCFDA fluorescence in young and aged subcutaneous ASCs. (C, D) Subcutaneous ASCs (C) and visceral (D) were isolated, treated with DCFDA dye ex vivo, and analyzed by FACS. Graphs are representative of eight mice per group and ASCs from two mice were pooled together to achieve 5000 events count. (n = 2 male and 2 female mice per group) P value < 0.05 = ^*, ns = non-significant.