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. 2022 Jun;10(12):683. doi: 10.21037/atm-22-2753

Figure 2.

Figure 2

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.