p53/p21 Pathway Involved in Mediating Cellular Senescence of Bone Marrow-Derived Mesenchymal Stem Cells from Systemic Lupus Erythematosus Patients
Figure 6
p21 knockdown reversed the ageing characteristics of BM-MSCs from SLE patients. (a) The morphology of BM-MSCs showed more spindle-shaped fibroblast-like growth after knockdown p21 when compared with BM-MSCs culture without si-p21. (b) Growth curve of BM-MSCs treated with and without si-p21 was tested by cell-counting assay. It has showed that, when p21 was knocked down, the cell proliferation rate was increased. Each point represents quantities relative to the BM-MSCs from SLE treated without si-p21 at Day 1 (*) (#). ((c)-(d)) CFU of BM-MSCs from treated with si-p21 were increased; each bar represents quantities relative to BM-MSCs from SLE treated without si-p21 and is mean ± SD of three experiments (#). ((e)-(f)) After treated with si-p21, the number of SA-β-gal-positive cells was obviously decreased; each bar represents quantities relative to BM-MSCs from SLE treated without si-p21 and is mean ± SD of three experiments (#). (g) DNA content between BM-MSCs culture with and without si-p21 from SLE patients was compared by flow-cytometry. (h) When BM-MSCs from SLE patients treated with si-p21, the percentage of cells in G0/G1 was obviously decreased, and increase in percentage of cells in S phases was seen in SLE patients. Each bar represents quantities relative to BM-MSCs from SLE treated without si-p21 and is mean ± SD of three experiments (*; #). ((i)–(k)) Expressions of cyclin E, CDK2, and p-Rb in BM-MSCs from SLE patients were tested by Western blot analyses and quantification analyses between culture with and without si-p21. The relative levels of protein expressions were normalized to GAPDH expression. Values are means ± SD of three experiments (*; #).
