Figure 1.

Strategies for the Discovery of Novel miRNA/Target Gene Sets and Confirmation of Changes of miR-27a-3p

(A) Strategy to find a novel miRNA and its target. Primarily cultured rat vascular smooth muscle cells (rVSMCs) were treated for 6 days and total RNA was prepared for a miRNA microarray and mRNA microarray. The samples were duplicated for each condition. The averaged value was used for analysis. (B) Gene tree analysis with the miRNA microarray results (GEO: GSE130486). (C) Changes in miR-27a-3p in microarray results in rVSMCs. Two values were averaged. (D) qRT-PCR analysis. Inorganic phosphate (Pi) treatment for 6 days significantly reduced the miR-27a-3p transcript amount in A10 cells. (E) Time course of Pi treatment and the decrease in miR-27a-3p amount. (F) The miR-27a-3p transcript level was significantly downregulated in the aorta obtained from vitamin D₃-administered mice. Error bars indicate SD. ∗p < 0.05, ∗∗p < 0.01.