Journal: Physiological Reports

Article Title: miR‐145 transgenic mice develop cardiopulmonary complications leading to postnatal death

doi: 10.14814/phy2.15013

Figure Lengend Snippet: Overexpression of miR ‐ 145 in the cardiovascular system leads to premature death in mice. Kaplan–Meier survival curve to illustrate the range in which miR ‐ 145 transgenic mice (miR ‐ 145Tg ; MCC) succumb. Genotype of control mice were +/+;+/+, miR ‐ 145Tg /+;+/+, or +/+; MCC /+ (n = 18) (a). LacZ expression demonstrates expression of the Myocardin ‐ Cre transgene (MCC) in the hearts and lungs of mice at embryonic day (E)16.5 (b), and in the lungs in postnatal day 14 mice (c) (n = 3)

Article Snippet: Human pulmonary artery smooth muscle cells (Lonza CC‐2581) were plated in a 12‐well plate at 5 × 10 4 cells/well for 12 h. Cells were transfected with miR‐145‐3p (Sigma assay HM10225), miR‐145‐5p (Ambion assay ID: MC11480) or control RNA mimic at 40 nM using Lipofectamine RNAiMAX (Invitrogen) as per manufacturer's instructions.

Techniques: Over Expression, Transgenic Assay, Expressing

Journal: Physiological Reports

Article Title: miR‐145 transgenic mice develop cardiopulmonary complications leading to postnatal death

doi: 10.14814/phy2.15013

Figure Lengend Snippet: miR ‐ 145 transgenic mice exhibit right ventricular hypertrophy. Dissected hearts from control and miR ‐ 145Tg ; MCC mice show miR ‐ 145 overexpressing mice have an enlarged right ventricle (arrow) (n = 7) (a). Fulton index show weight ratio of right ventricle (RV) to left ventricle and septum is significantly greater in the miR ‐ 145Tg ; MCC hearts compared to controls (b). H&E staining of cross‐sectioned hearts demonstrate increased thickness of the RV wall in miR ‐ 145Tg ; MCC mice (n = 7) (c). Genotype of control mice were +/+;+/+, miR ‐ 145Tg /+;+/+, or +/+; MCC /+ (n = 7)

Article Snippet: Human pulmonary artery smooth muscle cells (Lonza CC‐2581) were plated in a 12‐well plate at 5 × 10 4 cells/well for 12 h. Cells were transfected with miR‐145‐3p (Sigma assay HM10225), miR‐145‐5p (Ambion assay ID: MC11480) or control RNA mimic at 40 nM using Lipofectamine RNAiMAX (Invitrogen) as per manufacturer's instructions.

Techniques: Transgenic Assay, Staining

Journal: Physiological Reports

Article Title: miR‐145 transgenic mice develop cardiopulmonary complications leading to postnatal death

doi: 10.14814/phy2.15013

Figure Lengend Snippet: Lungs of miR ‐ 145Tg ; MCC mice exhibit alveolar remodeling and increased vessel muscularization. H&E staining of lung sections of control and miR‐145 transgenic mice show alterations in lung architecture and alveolar remodeling (a) (n = 10). Quantification of structural differences by mean linear intercept (MLI) indicates transgenic mice are significantly different from control mice (b). Degree of blood vessel muscularization was determined by measuring wall thickness (c), counting cell number (d), and staining for smooth muscle α‐actin (Acta2) (e), which shows an increase in smooth muscle surrounding small caliber arteries in the lung (n = 6)

Article Snippet: Human pulmonary artery smooth muscle cells (Lonza CC‐2581) were plated in a 12‐well plate at 5 × 10 4 cells/well for 12 h. Cells were transfected with miR‐145‐3p (Sigma assay HM10225), miR‐145‐5p (Ambion assay ID: MC11480) or control RNA mimic at 40 nM using Lipofectamine RNAiMAX (Invitrogen) as per manufacturer's instructions.

Techniques: Staining, Transgenic Assay

Journal: Physiological Reports

Article Title: miR‐145 transgenic mice develop cardiopulmonary complications leading to postnatal death

doi: 10.14814/phy2.15013

Figure Lengend Snippet: miR ‐ 145 induces the expression of smooth muscle marker genes in pulmonary artery smooth muscle cells. Human pulmonary artery smooth muscle cells (hPASMCs) were transfected with control (Con), miR ‐ 145 ‐ 3p (3p), or miR ‐ 145 ‐ 5p (5p) mimics followed by RNA isolation and qPCR to detect marker gene expression as indicated (n = 6). * p < 0.05, relative to control

Article Snippet: Human pulmonary artery smooth muscle cells (Lonza CC‐2581) were plated in a 12‐well plate at 5 × 10 4 cells/well for 12 h. Cells were transfected with miR‐145‐3p (Sigma assay HM10225), miR‐145‐5p (Ambion assay ID: MC11480) or control RNA mimic at 40 nM using Lipofectamine RNAiMAX (Invitrogen) as per manufacturer's instructions.

Techniques: Expressing, Marker, Transfection, Isolation

Journal: Physiological Reports

Article Title: miR‐145 transgenic mice develop cardiopulmonary complications leading to postnatal death

doi: 10.14814/phy2.15013

Figure Lengend Snippet: Single cell RNA‐seq of wild‐type and miR ‐ 145Tg P13 lungs. t‐SNE plot of cell clusters from wild‐type and miR ‐ 145 ‐ Tg ; MCC lungs combined (a). Overlap of cells from wild‐type (CTRL) and miR ‐ 145 ‐ Tg ; MCC (MIR145) lungs (b). Cell type identification of clusters (c) and violin plots of smooth muscle marker genes reveals vascular (cluster 10) and airway (cluster 15) smooth muscle cell subpopulations (d)

Article Snippet: Human pulmonary artery smooth muscle cells (Lonza CC‐2581) were plated in a 12‐well plate at 5 × 10 4 cells/well for 12 h. Cells were transfected with miR‐145‐3p (Sigma assay HM10225), miR‐145‐5p (Ambion assay ID: MC11480) or control RNA mimic at 40 nM using Lipofectamine RNAiMAX (Invitrogen) as per manufacturer's instructions.

Techniques: RNA Sequencing Assay, Marker