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 6. Rmst modulates Notch signaling pathway by neutralizing a panel of Notch-targeting miRNAs in BMP9-induced osteogenic differentiation. (A) Silencing Rmst reduces the expression of most Notch receptors and ligands. Exponentially growing iMADs were infected with Ad-GFP and Ad-simRmst for 72h. Total RNA was isolated and subjected to qPCR analysis use primers for the indicated genes. Each qPCR assay condition was done in triplicate. Gapdh was used as a reference gene. “*”, p<0.05, “**”, p<0.01, AdR-simRmst group vs. Ad-GFP group. (B) Putative target sites on Rmst for several Notch-targeting miRNAs. (C) BMP9 suppresses the expression of Notch-targeting miRNAs in MSCs. The iMADs were infected with Ad-GFP or Ad-BMP9 for 72h. Total RNA was isolated and subjected to TqPCR analysis. Each qPCR assay condition was done in triplicate. Gapdh was used as a reference gene. “*”, p<0.05, “**”, p<0.01, when Ad-BMP9 group vs. Ad-GFP group. (D) Silencing Rmst restores the expression of several Notch-targeting miRNAs in MSCs. The iMADs cells were infected with Ad-GFP or AdR-simRmst for 72h. Total RNA was isolated and subjected to TqPCR analysis. Each qPCR assay condition was done in triplicate. Gapdh was used as a reference gene. “*”, p<0.05, “**”, p<0.01, when AdR-simRmst group vs. Ad-GFP group.