plasmid dna  (OriGene)

Journal: PLoS ONE

Article Title: Direct Reprogramming of Human Fibroblasts to Hepatocyte-Like Cells by Synthetic Modified mRNAs

doi: 10.1371/journal.pone.0100134

Figure Lengend Snippet: Production and transfection of synthetic modified mRNAs. (A) Scheme for producing DNA templates with synthetic UTR and PolyA sequences attached to the ORFs of interest. Primers used in the ORF PCR are gene specific. Primers used in template-tail PCR are independent of the gene and are always the same. Tailed-templates can be generated, purified, and used for overnight in vitro transcription in under two hours of bench time using this method. (B–I) Concentration-dependent translation and proper localization of GFP and nuclear GFP mmRNA at 80 ng, 200 ng, 500 ng, and 1000 ng per well in a 12-well plate. Nuclei are stained blue by Hoechst.

Article Snippet: Open reading frame (ORF) PCR amplifications of DNA encoding C/EBPA FOXA1, FOXA2, FOXA3, GATA4, GATA6, HHEX, HNF1A, HNF1B, HNF4A, and HNF6Awere templated from DNA plasmids containing each of the respective human ORFs (Origene, Rockville, MD).

Techniques: Transfection, Modification, Polymerase Chain Reaction, Generated, Purification, In Vitro, Concentration Assay, Staining

Journal: BMC Cell Biology

Article Title: Restoration of angiogenic capacity of diabetes-insulted mesenchymal stem cells by oxytocin

doi: 10.1186/1471-2121-14-38

Figure Lengend Snippet: Reduced KLF2 level of MSC from DM bone marrow was restored by oxytocin. (A) KLF2 mRNA was assessed in DM-MSC treated with various concentrations oxytocin (n=3). (B) KLF2 mRNA was analyzed in MSC from non-DM or DM bone marrow treated with PBS or oxytocin for 24 hours (n=3). (C) The level of KLF2 protein was reduced in DM-MSC, whereas restored by oxytocin (n=3). (D) Oxytocin-induced KLF2 mRNA was assessed in DM-MSC transfected with scrambled siRNA or KLF2 siRNA (n=3). (E) After transfection with KLF2 plasmid for 24 hours, 293T cells were treated with PBS or oxytocin for 24 hours. KLF2 protein level was examined by Western blot (n=3). Quantification analysis was performed and data in the graph represented the mean ± standard deviation.

Article Snippet: Plasmid DNA or siRNA transfection KLF2 plasmid DNA was purchased from OriGene Technologies (Rockville, USA).

Techniques: Transfection, Plasmid Preparation, Western Blot, Standard Deviation

Journal: International Journal of Biological Sciences

Article Title: Bisdemethoxycurcumin Enhances the Sensitivity of Non-small Cell Lung Cancer Cells to Icotinib via Dual Induction of Autophagy and Apoptosis

doi: 10.7150/ijbs.40042

Figure Lengend Snippet: Combination of icotinib and BDMC suppresses EGFR activity by downregulating expressions of SP1 and HDAC1/HDAC2. ( A ) H460 and H1781 cells were pre-treated with icotinib and/or BDMC at the doses indicated for 6 h, and then stimulated with EGF at indicated dose for 1 h. Then cell lysates were prepared and subjected to immunoblot assay to detect the protein expressions as indicated. ( B ) Expressions of Sp1/Sp3 and other indicated proteins in control and SP1or SP3 knocked down cells were detected by immunoblot assay in H460 and H1781 cells. ( C ) H460 and H1781 cells were transfected with Sp1 plasmids (PCMV6-Sp1) and control plasmids DNA (PCMV6). After 8 h, cells were treated with BDMC at indicated doses for an additional 48 h. Then the expressions of Sp1 and HDAC1/HDAC2 proteins were detected by immunoblot assay. ( D and E ) Before and after transfection with or without siSp1 or PCMV6-Sp1, H460 and H1781 cells were treated with icotinib (5 µM) plus BDMC (10 µM) for 48 h, and HDCA1 mRNA expression was analyzed by real-time quantitative-PCR, as described in Materials and Methods. * P

Article Snippet: Plasmid transient transfection The Sp1 and control plasmid DNA (PCMV6 XL8) used in this study were purchased from OriGene (Rockville, MD).

Techniques: Activity Assay, Transfection, Expressing, Real-time Polymerase Chain Reaction

Journal: Journal of Cardiothoracic Surgery

Article Title: Variation of perioperative plasma mitochondrial DNA correlate with peak inflammatory cytokines caused by cardiac surgery with cardiopulmonary bypass

doi: 10.1186/s13019-015-0298-6

Figure Lengend Snippet: DNA isolation and rt-PCR for mtDNA

Article Snippet: Plasmid DNA with complementary DNA sequence for human mtDNA was obtained from ORIGENE (SC101172, USA).

Techniques: DNA Extraction, Reverse Transcription Polymerase Chain Reaction