dlr detection kit  (Promega)

Journal: American Journal of Translational Research

Article Title: miR-370 impacts the biological behavior of lung cancer cells by targeting the SMAD1 signaling pathway

doi:

Figure Lengend Snippet: Targeting association between miR-370 and SMAD1. A. Prediction of targets of miR-370 by Targetscan and miRDB. B. Analysis of SMAD1 expression in NSCLC by TCGA and GTEx. C. Detection of SMAD1 expression in LC cell lines by RT-qPCR and western blot. D. Analysis of the targeting association between miR-370 and SMAD1 by DLR assay. Note: miR: MicroRNA; LC: Lung Cancer; RT-qPCR: Real-time Fluorescence Quantitative Polymerase Chain Reaction; SMAD1: SMAD Family Member 1; DLR: Dual Luciferase Reporter; WT: Wild Type; MUT: Mutant; WB: western blot, ***P<0.001, ****P<0.0001.

Article Snippet: After 48 h of transfection, the luciferase activity was determined through a DLR gene detection kit (Promega, Madison, WI, the States) following the corresponding protocol.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Fluorescence, Real-time Polymerase Chain Reaction, Luciferase, Mutagenesis

Journal: Frontiers in Pediatrics

Article Title: Long Non-coding RNA TUG1 Modulates Expression of Elastin to Relieve Bronchopulmonary Dysplasia via Sponging miR-29a-3p

doi: 10.3389/fped.2020.573099

Figure Lengend Snippet: miR-29a-3p is a target gene of TUG1 and inversely regulated by TUG1. (A) The binding sites between TUG1 and miR-29a-3p were predicted by starbase2.0. (B) Relative luciferase activity of TUG1 vector was detected by dual-luciferase reporter (DLR) assay. ** P < 0.01 vs. miR-negative control (NC). (C) After transfection of miR-29a-3p mimics, relative expression of miR-29a-3p was detected by quantitative real-time polymerase chain reaction (qRT-PCR) in hyperoxia-induced MLE-12 cells. ** P < 0.01 vs. miR-NC. (D) After transfection of TUG1, relative expression of miR-29a-3p was detected by qRT-PCR in hyperoxia-induced MLE-12 cells. ** P < 0.01 vs. miR-NC. (E) Relative expression of miR-29a-3p was detected by qRT-PCR in lung tissues of BPD mice. P < 0.001 vs. Blank. Each experiment was repeated three times.

Article Snippet: The above vectors were transfected into MLE-12 cells, together with miR-NC/mmu-miR-29a-3p (RiboBio) and miR-NC/miR-29a-3p mimics (RiboBio) for 48 h. DLR assay kit (Biovision, Milpitas, CA, USA) was used for detection of relative luciferase activity.

Techniques: Binding Assay, Luciferase, Activity Assay, Plasmid Preparation, Negative Control, Transfection, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Frontiers in Pediatrics

Article Title: Long Non-coding RNA TUG1 Modulates Expression of Elastin to Relieve Bronchopulmonary Dysplasia via Sponging miR-29a-3p

doi: 10.3389/fped.2020.573099

Figure Lengend Snippet: Elastin (ELN) is a downstream target of miR-29a-3p and negatively correlated with miR-29a-3p. (A) The binding sites between miR-29a-3p and ELN were predicted by starbase2.0. (B) Relative luciferase activity of ELN vector was detected by DLR assay. ** P < 0.01 vs. miR-negative control (NC). (C) After transfection with miR-29a-3p mimics, expression of ELN was detected by Western blot in hyperoxia-induced MLE-12 cells. ** P < 0.01 vs. miR-NC. (D) Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect relative expression of ELN in lung tissues of BPD mice. P < 0.001 vs. Blank. Each experiment was repeated three times.

Article Snippet: The above vectors were transfected into MLE-12 cells, together with miR-NC/mmu-miR-29a-3p (RiboBio) and miR-NC/miR-29a-3p mimics (RiboBio) for 48 h. DLR assay kit (Biovision, Milpitas, CA, USA) was used for detection of relative luciferase activity.

Techniques: Binding Assay, Luciferase, Activity Assay, Plasmid Preparation, Negative Control, Transfection, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR