Research Paper Volume 8, Issue 9 pp 2006—2022

Figure 5. SATB2 regulated age-related properties of AB-BMSCs by Nanog pathway. (A, B) Real-time PCR and western blot results showed the decrease of Nanog, SOX2 and OCT4 expressions after Nanog knockdown in AB-BMSCs with SATB2 overexpression (Lev-Satb2/Si-Nanog) in relative to control (Lev-Satb2/Scramble). (C) Immunofluorescence staining verified Nanog reduction upon Nanog knockdown mediated by siRNA. (D, E) Nanog knockdown increased the number of SA-β-Gal positive cells and P16, P21 and P53 abundance in AB-BMSCs with SATB2 overexpression. (F, G) Nanog knockdown largely abrogated the effects of SATB2 overexpression in AB-BMSCs from older donors as demonstrated by weaker Alizarin red staining and reduced RUNX2, OSX and OPN expression. *p< 0.05, ***p< 0.001. Scale bar = 100 μm.