Journal: Human Genomics

Article Title: Downregulation of miR-542-3p promotes osteogenic transition of vascular smooth muscle cells in the aging rat by targeting BMP7

doi: 10.1186/s40246-019-0245-z

Figure Lengend Snippet: miR-542-3p targeted BMP7. a After 7 days of induction by β-GP, the protein expression of BMP7 was detected by Western blot. yVSMCs overexpressing miR-542-3p (miR-542-3p) expressed an obviously lower level of BMP7 than control yVSMCs and yVSMCs infected with an empty virus vector. b Quantification of BMP7 expression by densitometric analysis. The relative expression of BMP7 was normalized to that of GADPH. n = 3. Versus control and vector group, ** p < 0.01. c BMP7 3′-UTR constructs used in luciferase assay. d Effect of miR-542-3p overexpression on a dual luciferase reporter plasmid containing the BMP7 3′-UTR was analyzed. HEK293T cells were co-transfected with either the BMP7 WT or BMP7 MUT or an empty vector and miR-542-3p (miR-542-3p). Renilla luciferase activity was quantified and normalized to firefly luciferase activity. n = 3. Versus vector group, ** p < 0.01

Article Snippet: The 3′-UTR of the luciferase reporter containing the BMP7 WT sequence or the BMP7 MUT sequence was inserted into the pmirGLO vector (Promega, Beijing, China).

Techniques: Expressing, Western Blot, Infection, Plasmid Preparation, Construct, Luciferase, Over Expression, Transfection, Activity Assay

Journal: Human Genomics

Article Title: Downregulation of miR-542-3p promotes osteogenic transition of vascular smooth muscle cells in the aging rat by targeting BMP7

doi: 10.1186/s40246-019-0245-z

Figure Lengend Snippet: miR-542-3p regulated osteogenic transition by targeting BMP7. yVSMCs overexpressing miR-542-3p (miR-yVSMCs) were transduced with BMP7-overexpressing lentivirus or control empty virus. After 7 days of induction by β-GP, the mRNA ( a ) and protein ( b ) expression levels of smooth muscle markers (SM22α and calponin) and osteogenic markers (OPN and Runx2) were detected. c Quantification of protein expression levels by densitometric analysis. d Calcium deposition detected by Alizarin red staining. e Quantification of calcium deposition. Control: miR-yVSMCs; Vector: miR-yVSMCs transduced with control empty virus; BMP7 overexpression: miR-yVSMCs transduced with BMP7-overexpressing lentivirus. n = 3. Versus control and vector groups, * p < 0.05, ** p < 0.01

Article Snippet: The 3′-UTR of the luciferase reporter containing the BMP7 WT sequence or the BMP7 MUT sequence was inserted into the pmirGLO vector (Promega, Beijing, China).

Techniques: Transduction, Expressing, Staining, Plasmid Preparation, Over Expression

Journal: Human Genomics

Article Title: Downregulation of miR-542-3p promotes osteogenic transition of vascular smooth muscle cells in the aging rat by targeting BMP7

doi: 10.1186/s40246-019-0245-z

Figure Lengend Snippet: A proposed schematic diagram of miR-542-3p regulating the osteogenic transition of VSMCs in aging rats. MiR-542-3p expression is downregulated in oVSMCs. As a target of MiR-542-3p, the expression of BMP7 is upregulated which results in the osteogenic differentiation of VSMCs, suggesting the role of miR-542-3p in the vascular osteogenic transition in aging rat

Article Snippet: The 3′-UTR of the luciferase reporter containing the BMP7 WT sequence or the BMP7 MUT sequence was inserted into the pmirGLO vector (Promega, Beijing, China).

Techniques: Expressing

Journal: Oncology Letters

Article Title: miR-224 enhances invasion and metastasis by targeting HOXD10 in non-small cell lung cancer cells

doi: 10.3892/ol.2018.8245

Figure Lengend Snippet: miR-224 downregulates HOXD10 expression by targeting its 3′UTR. (A) An illustration of WT and MUT 3′-UTR of HOXD10 showing a putative miR-224 target site. (B) The luciferase activities of HOXD10-WT, and HOXD10-MUT in H358 and A549 cells transfected with miR-224 mimic. (C) HOXD10 mRNA level was remarkably lower examined by qRT-PCR and it was inversely correlated with miR-224 level in NSCLC tissues (20 paired tissues). (D) Protein level of HOXD10 and GAPDH in H358 and A549 cells. **P<0.01, ***P<0.001.

Article Snippet: The pmirGLO vectors with wild-type (WT) or mutant type (MUT) miR-224 binding site in HOXD10 3′-UTR were constructed by GenePharma Co., Ltd. (Shanghai, China).

Techniques: Expressing, Luciferase, Transfection, Quantitative RT-PCR