Research Paper Volume 11, Issue 24 pp 12476—12496

lncRNA Rmst acts as an important mediator of BMP9-induced osteogenic differentiation of mesenchymal stem cells (MSCs) by antagonizing Notch-targeting microRNAs

Figure 1. BMP9-induced expression of lncRNA Rmst and construction of adenoviral vector-mediated siRNA knockdown of Rmst expression in MSCs. (A) BMP9 induces the expression of lncRNA Rmst in MSCs. Subconfluent iMADs were infected with Ad-GFP or Ad-BMP9. At the indicated time points, total RNA was isolated and subjected to quantitative TqPCR analysis of Rmst expression. Gapdh was used as a reference gene. “**” p<0.001 when compared with Ad-GFP control group. Each assay condition was done in triplicate. (B) The transcriptomic arrangement of mouse lncRNA Rmst and the locations and sequences of three siRNA targeting sites are shown. (C) A recombinant adenoviral vector, called AdR-simRmst expressing the three siRNA sites, was constructed. To assess the Rmst knockdown efficiency, subconfluent iMADs were infected with AdR-simRmst or control Ad-GFP. At the indicated time point, total RNA was isolated and subjected to quantitative TqPCR analysis of Rmst expression. Gapdh was used as a reference gene. “**” p<0.001 when compared with Ad-GFP control group. Each assay condition was done in triplicate.