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. 2022 Aug 3;12:13361. doi: 10.1038/s41598-022-17579-0

Figure 2.

Figure 2

miR-101 targets Ezh2 and stimulates mineral deposition in differentiating MC3T3 cells. Experimental set-up of MC3T3 transfection and differentiation (A). Cells were plated and transfected in growth medium. Cells were then maintained in growth medium (GM) and differentiation medium (DM) for indicated times. Media were changed every two to three days (Micro. Microscopy, WB Western blotting, AR stain alizarin red staining). Representative light and fluorescence microscopy images of Cy3 dye-labeled pre-miRNA negative control transfected MC3T3 cells one day after transfection (B). Representative live (green) and dead (red) fluorescence microscopy images of Cy3 negative control and miR-101a-3p transfected MC3T3 cells (C). Western blot analysis of MC3T3 cell lysates transfected with Cy3 negative control and miR-101-3p (D). Alizarin red staining (E) and quantification (F) of differentiating MC3T3 cells (day 28) transfected with CY3 control, miR-101a-3p, and miR-27b [n = 3, mean ± standard deviation (STD)]. *p < 0.05 and ***p < 0.001. All experiments were repeated at least three times.