Journal: Biological Research

Article Title: DNMT1-mediated PPARα methylation aggravates damage of retinal tissues in diabetic retinopathy mice

doi: 10.1186/s40659-021-00347-1

Figure Lengend Snippet: DNMT1 overexpression reversed the impact of PPARα overexpression on apoptosis and ROS levels in HG-treated HRCPs. HRCPs were co-transfected with pCEP4-PPARα, pCEP4-NC and pcDNA3.1-DNMT1 or pcDNA3.1-NC, and then treated with 30 mM glucose. QRT-PCR ( A ) and WB ( B , C ) were performed to assess PPARα gene and protein expression in HRCPs. D , E Flow cytometry was performed to estimate apoptosis of HRCPs. F , G The levels of ROS in HRCPs was detected by DCFH-DA fluorescent probe. * P < 0.05 vs. Vector1; # P < 0.05 vs. PPARα + vector2

Article Snippet: The vector pcDNA3.1-DNMT1 overexpressed DNMT1, and the corresponding empty vector pcDNA3.1-NC (vector2) were constructed by GeneChem.

Techniques: Over Expression, Transfection, Quantitative RT-PCR, Expressing, Flow Cytometry

Journal: World Journal of Surgical Oncology

Article Title: Rs6757 in microRNA-3976 binding site of CD147 confers risk of hepatocellular carcinoma in South Chinese population

doi: 10.1186/s12957-022-02724-w

Figure Lengend Snippet: Sequencing results of the inserts in CD147-TT-pSICHECK2 plasmid. Arrows indicate the inserted rs6757 TT genotype fragment

Article Snippet: The rs6757 TT genotype inserts were digested by XhoI/NotI , gel-purified, extracted, and then subcloned into the XhoI/NotI site of pSICHECK2 vector (Promega).

Techniques: Sequencing, Plasmid Preparation

Journal: World Journal of Surgical Oncology

Article Title: Rs6757 in microRNA-3976 binding site of CD147 confers risk of hepatocellular carcinoma in South Chinese population

doi: 10.1186/s12957-022-02724-w

Figure Lengend Snippet: The Blast of the inserts in CD147-TT-pSICHECK2 plasmid. The sequencing results were analyzed by Blast, the inserted sequence of target gene was 100% consistent with the sequence of CD147 rs6757 TT genotype in the NCBI database

Article Snippet: The rs6757 TT genotype inserts were digested by XhoI/NotI , gel-purified, extracted, and then subcloned into the XhoI/NotI site of pSICHECK2 vector (Promega).

Techniques: Plasmid Preparation, Sequencing

Journal: World Journal of Surgical Oncology

Article Title: Rs6757 in microRNA-3976 binding site of CD147 confers risk of hepatocellular carcinoma in South Chinese population

doi: 10.1186/s12957-022-02724-w

Figure Lengend Snippet: Sequencing results of the inserts in CD147-CC-pSICHECK2 plasmid. The sequencing result showed that the mutation from nucleotide T to C at the target site (marked in green) of CD147 gene had successfully achieved

Article Snippet: The rs6757 TT genotype inserts were digested by XhoI/NotI , gel-purified, extracted, and then subcloned into the XhoI/NotI site of pSICHECK2 vector (Promega).

Techniques: Sequencing, Plasmid Preparation, Mutagenesis

Journal: World Journal of Surgical Oncology

Article Title: Rs6757 in microRNA-3976 binding site of CD147 confers risk of hepatocellular carcinoma in South Chinese population

doi: 10.1186/s12957-022-02724-w

Figure Lengend Snippet: The Blast of the inserts in CD147-CC-pSICHECK2 plasmid. After mutating from nucleotide T to C, the inserted sequence was 99% consistent with the sequence of CD147 in the NCBI database by Blast analysis

Article Snippet: The rs6757 TT genotype inserts were digested by XhoI/NotI , gel-purified, extracted, and then subcloned into the XhoI/NotI site of pSICHECK2 vector (Promega).

Techniques: Plasmid Preparation, Sequencing

Journal: World Journal of Surgical Oncology

Article Title: Rs6757 in microRNA-3976 binding site of CD147 confers risk of hepatocellular carcinoma in South Chinese population

doi: 10.1186/s12957-022-02724-w

Figure Lengend Snippet: The R/F analysis of a luciferase reporter vector. A Luciferase expression was significantly reduced following transfection with CD147-TT-pSICHECK2 ( P < 0.01); the translational suppression of miR-3976 can be reversed by its inhibitor. B There was no significant statistical difference between the groups following transfection with CD147-CC-pSICHECK2. C Luciferase activity was decreased by miR-3976 in dose-dependent manner for the constructs with a TT genotype but not changed for constructs with a CC genotype at the rs6757:T>C polymorphism. Each transfection was carried out in triplicate. * denotes a p value < 0.05; ** denotes a p value < 0.01

Article Snippet: The rs6757 TT genotype inserts were digested by XhoI/NotI , gel-purified, extracted, and then subcloned into the XhoI/NotI site of pSICHECK2 vector (Promega).

Techniques: Luciferase, Plasmid Preparation, Expressing, Transfection, Activity Assay, Construct

Journal:

Article Title: Neuroendocrine differentiation factor, IA-1, is a transcriptional repressor and contains a specific DNA-binding domain: identification of consensus IA-1 binding sequence

doi:

Figure Lengend Snippet: IA-1 zinc-fingers 2 and 3 are essential for transcriptional activity. Various zinc-finger constructs were fused in frame with hEgr-1 AD (a.a. 1–147). Co-transfection of zinc-finger mutants with the IA-1 –426/+40 bp promoter/pCAT3 reporter gene into β-TC-1 cells revealed that zinc-fingers 2 and 3 are the key motifs that contribute to the transcriptional activity. Zinc-finger 3 alone exhibits 50% of the control activity, whereas the combination of zinc-finger 3 and 4 only exhibits 25% of the control activity. The data are expressed as fold increase over the empty pcDNA3 expression vector. A CMV–βgal vector is used to normalize transfection efficiency. The graph represents the average of three separate experiments and SEM.

Article Snippet: The –426/+40 bp IA-1 promoter/CAT construct was generated by subcloning a Nhe I/ Xho I –426/+40 bp IA-1 promoter fragment into the Nhe I/ Xho I site of the pCAT3 vector (Promega).

Techniques: Zinc-Fingers, Activity Assay, Construct, Cotransfection, Expressing, Plasmid Preparation, Transfection

Journal: Oncotarget

Article Title: microRNA-203 suppresses invasion and epithelial-mesenchymal transition induction via targeting NUAK1 in head and neck cancer

doi: 10.18632/oncotarget.6972

Figure Lengend Snippet: A. Luciferase assays were performed with pmirGLO vector containing the 3′-UTR of NUAK1 and the graph shows the relative luciferase activity in mature miR-203- or control miRNA-transfected cells. The data were normalized to control samples and results are presented as means ± SD. ** P < 0.01. B. NUAK1 expression was examined in control- or mature miR-203-transfected HNSCC cells (KOSCC25B, SpSCC, and HOC313) by western blotting analysis. The graph shows the NUAK1/β-actin ratio by densitometric analysis. C. NUAK1 expression was examined in control- or miR-203 inhibitor-transfected HSC2 cells by western blotting analysis. The graph shows the NUAK1/β-actin ratio by densitometric analysis.

Article Snippet: The oligonucleotides had the following sequences: SNAI2-1: 5′-CATTGCTGCCAAATCATTTCAA-3′ SNAI2-2: 5′-TTACTATTTTAAAACATTTTAA-3′ SNAI2-3: 5′-TAATGTACTTAAACTATTTCAA-3′ The full-length 3′-UTR of human NUAK1 was amplified by PCR ( ) from genomic DNA and cloned at the EcoRI site into the pmirGLO vector (Promega).

Techniques: Luciferase, Plasmid Preparation, Activity Assay, Transfection, Expressing, Western Blot