Journal: Journal of Orthopaedic Translation
Article Title: De-osteogenic-differentiated mesenchymal stem cells accelerate fracture healing by mir-92b
doi: 10.1016/j.jot.2020.10.009
Figure Lengend Snippet: Mir-92b promoted osteogenesis in MSCs (A–B) The top three up/down-regulated microRNAs in De-Os-MSCs were listed, and verified by qPCR (B) (C–E) The scrambled control, let-7e, mir-10b, mir-20a, mir-92b, mir-371 and mir-373 were transduced into MSCs with lentiviruses. The overexpression of each microRNA was verified by qPCR (C). The infected MSCs were induced to undergo osteogenic differentiation for 10 days, then the calcium deposits were stained with Alizarin Red S (D), and quantified (E) (F) Total RNA was extracted from MSCs infected with mir-92b or scrambled control. The mRNA expression levels of Osterix, Runx2, OPN and ALP were detected by qPCR. β-actin was used as an internal control. The data was expressed as mean ± SD (n = 3). ∗p < 0.05 (G–I) Total proteins were extracted from MSCs transduced with scrambled control or mir-92b. Then the proteins were analyzed by western blot using indicated antibodies. The protein levels of pERK (H) and pJNK (I) was normalized to ERK and JNK1 respectively. All the data represent mean ± SD of three independent experiments. ∗p < 0.05 (J–K) The mir-92b antagmir was transfected into MSCs, then the cells were treated with osteogenic induction medium for 10 days, the calcium deposits were stained with Alizarin Red S (J), the changes of osteogenesis-related genes was checked by qPCR (K).
Article Snippet: The microRNA microarray analysis was performed by the Annoroad Gene Technology Corporation (Beijing, China).
Techniques: Control, Over Expression, Infection, Staining, Expressing, Transduction, Western Blot, Transfection