Journal: Annals of Translational Medicine
Article Title: MiR-20a: a mechanosensitive microRNA that regulates fluid shear stress-mediated osteogenic differentiation via the BMP2 signaling pathway by targeting BAMBI and SMAD6
doi: 10.21037/atm-22-2753
Figure Lengend Snippet: Overexpression of miR-20a activates osteogenic MC3T3-E1 cell differentiation. After transfecting the cells with miR-20a double-stranded miRNA mimics (50 or 100 nM), miRNA NC (50 or 100 nM), or transfection reagent only (control) for 12 h, the cultures of confluent MC3T3-E1 cells were placed in α-MEM medium with β-glycerophosphate (10 mM) and 50 µg/mL ascorbic acid. (A) qRT-PCR assays for ALP mRNA at 24 h. (B) Detection of ALP activity on day 3. (C) Determination of ECM mineralization by stained with Alizarin red S on day 14. Relative means and standard deviations were shown underneath (P<0.001; scale bar: 50 µm). Data are expressed as mean ± SD for n=3; **, P<0.01 vs. NC; ***, P<0.001 vs. NC. NC, negative control; qRT-PCR, real-time quantitative polymerase chain reaction; ALP, alkaline phosphatase; ECM, extracellular matrix.
Article Snippet: Cell culture Culturing of MC3T3-E1 cells, a murine pre-osteoblastic cell line (ATCC, Manassas, VA, USA) was performed in minimum essential medium alpha medium (α-MEM) media (Life Technologies, Grand Island, NY, USA) with 1% penicillin/streptomycin (Life Technologies) and fetal bovine serum (FBS; 10%, Life Technologies).
Techniques: Over Expression, Cell Differentiation, Transfection, Control, Quantitative RT-PCR, Activity Assay, Staining, Negative Control, Real-time Polymerase Chain Reaction