Research Paper Volume 11, Issue 20 pp 9167—9187

Bone marrow-derived mesenchymal stem cells in three-dimensional co-culture attenuate degeneration of nucleus pulposus cells

Figure 5. Effects of ZMPSTE24 on senescence associated TGFβ/Smad signaling pathway. (A) Elisa result of TGFβ1 in senescent NPCs culture supernatant. (BC) Protein expressions of Smad2, Smad3, Smad4 and Smad5 under TNF-a inducing were visualized by western blot and quantified by Image J. n=3. (D) Protein expression of Smad2, Smad3, Smad4 and Smad5 were visualized by western blot and quantified by Image J. n=3. (EF) Spider-β-gal staining of NP cells between different neutralizing groups(Neut-TGFb and Neut-BMPR2). Combined with DAPI staining for nuclei. n=5, Scale bar, 20um. (GH) Protein expression of Smad4, Smad5 and p-Smad1/5/8 in NP cells with stable scramble vector (SC) or knockout of ZMPSTE24 were visualized by western blot and quantified by Image J. n=3. (IJ) Protein expression of Smad4, Smad5 and p-Smad1/5/8 in NP cells with or without BMSCs coculture were visualized by western blot and quantified by Image J. n=3. (KL) Spider-β-gal staining of NP cells between different coculture groups(NPCs or BMSCs) with or without lopinavir. Combined with DAPI staining for nuclei. n=5, Scale bar, 20um. Values represent means±S.D. Significant difference between different groups is indicated as *P < 0.05. PF: per field.