Figure 6: MiRNA expression changes caused by enamel matrix protein (EMD) exposure and EMD intake assay in presence of miR-153 overloading. | Scientific Reports

Figure 6: MiRNA expression changes caused by enamel matrix protein (EMD) exposure and EMD intake assay in presence of miR-153 overloading.

From: MiR-153 Regulates Amelogenesis by Targeting Endocytotic and Endosomal/lysosomal Pathways–Novel Insight into the Origins of Enamel Pathologies

Figure 6

There were two objectives of this experiment one was to evaluate the effect of miR-153 overloading on other miRNAs expressions after EMD exposure, and the second was to semi-quantify the effect of miR-153 overloading on EMD uptake. In both ALCs (a) and LS8 cells (b), treatment with EMD elicited significant downregulation of miR-3085, miR-298, miR-138, miR-135a and miR-376b. The ranges of miRNA expression fold changes in ALC cells were from ~−2.4 to −1.2 (b), and the fold changes of miRNAs in LS8 cells varied from ~−6.1 to −1.4 (a). The expression of miR-203 did not show any statistically significant changes in response to EMD uptake in either cell models (a,b), and decreased expression of miR-346 was only significant in ALCs (a). (c) After 6 h exposure to EMD, protein samples remaining in the medium of the cultured ALCs and LS8 cells were extracted for electrophoresis and silver staining. The presumable amelogenin bands are at ~20 kD. Positive control (Pos): no cells were seeded and EMD stock solution (2 mg/μl) was diluted to 500 μg/ml with FBS free culture medium; Negative control (Neg): enamel cells were seeded with no EMD treatment; MiR-153 transfection (miR): cells were transfected with miR-153 mimics and received treatment of EMD at 500 μg/ml for 6 h; Mock transfection with scrambled siRNA (Scrambled): cells were transfected with scrambled siRNAs and were exposed to 500 μg/ml EMD in the culture medium for 6 h. (d) 20 kD band densities were quantified relative to cell-free Positive control group. In ALCs and LS8 cells, miR-153 affected group showed significantly decreased relative band intensities compared with positive control group (ALC: ~−1.3-fold change positive/miR-153; LS8: ~−1.5-fold change positive/miR-153). When the miR-153 transfection group was compared with mock transfection group, the differences in relative band densities were determined to reach statistical significance only in ALCs (~−1.2-fold change scrambled/miR-153). *P < 0.05. NS Not significant.

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