Journal: World Journal of Stem Cells
Article Title: Silencing of Jumonji domain-containing 1C inhibits the osteogenic differentiation of bone marrow mesenchymal stem cells via nuclear factor-κB signaling
doi: 10.4252/wjsc.v16.i2.151
Figure Lengend Snippet: Osteogenic and adipogenic differentiation of bone mesenchymal stem cells. A: Bone mesenchymal stem cells (BMSCs) were isolated from mice bone marrow tissues. Flow cytometry was used to detect the expressions of CD29, CD31, CD90, CD45, CD34, MHCII, SCA-1 and CD11b; B-D: BMSCs were induced towards osteogenic differentiation. ALP staining was used to evaluate alkaline phosphatase (ALP) activity (B); calcium deposits were visualized by Alizarin red staining (C); the mRNA levels of osteogenic genes, including Osteocalin, Runt-related transcription factor 2 and ALP, were detected by reverse transcription coupled to the quantitative polymerase chain reaction (RT-PCR) (D); E and F: BMSCs were induced towards adipogenic differentiation. Representative images of Oil red O staining (E), the mRNA levels of adipogenic genes, such as peroxisome proliferator-activated receptor gamma and CCAAT enhancer-binding protein alpha, were measured by RT-PCR (F). All values are shown as mean ± SD. d P < 0.0001. n = 3. BMSC: Bone mesenchymal stem cell; ALP: Alkaline phosphatase; RunX2: Runt-related transcription factor 2; CEBPα: CCAAT enhancer-binding protein alpha; PPARγ: Peroxisome proliferator-activated receptor gamma.
Article Snippet: After centrifugation at 1500 rpm, the cells were incubated for 0.5 h with primary antibodies against CD29, CD31, CD34, CD45, CD90, MHCII, SCA-1 and CD11b (Biolegend, California, San Diego, United States and Becton Dickinson, Franklin Lakes, NJ, United States).
Techniques: Isolation, Flow Cytometry, Staining, Activity Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Binding Assay