Research Paper Volume 12, Issue 13 pp 12609—12621
Obesity is associated with senescence of mesenchymal stromal cells derived from bone marrow, subcutaneous and visceral fat of young mice
Figure 3. Apoptosis and senescence in MSCs from obese and control animals. (A) Representative FACS analysis of MSC apoptosis. The assay identifies early (Annexin V + and 7ADD −) and late apoptosis (Annexin V + and 7ADD +). Apoptosis is a continuous process and we calculated the percentage of apoptosis as the sum of early and late apoptotic cells. The histogram shows the mean percentage of Annexin V-positive cells. Data are expressed with standard deviation (n=6 animals for each experimental condition). (B) The graph shows mean percentage value of senescent cells determined by SPiDER-ßgal assay. Data are expressed with SD (n=6 animals for each experimental condition) **p<0.01, ***p<0.001. (C) The graph shows mean percentage value of cells showing discrete amount of intracellular ROS that are above the threshold detected by H2DCFDA assay. Data are expressed with SD (n=6 animals for each experimental condition) **p<0.01, ***p<0.001. (D) Western blot analysis of proteins regulating senescence. The picture shows a representative blot analysis the expression levels of Rb, p53, p21, p16/INK4, Gapdh (loading control). The graph shows mean expression levels (±SD, n = 6 biological replicates, *p<0.05).