cmv early enhancer promoter ebna 1 based pcep4 plasmid vector  (Thermo Fisher)

Journal: BMC Cancer

Article Title: Intratumoral delivery of IL-18 naked DNA induces T-cell activation and Th1 response in a mouse hepatic cancer model

doi: 10.1186/1471-2407-7-87

Figure Lengend Snippet: Immune response is enhanced by a single injection of naked IL-18 plasmid DNA . Each group (consisting of 4 mice) was administered an intratumoral injection of pCEP4 or naked mIL-18 DNA. Spleens of CT26 tumor-bearing mice treated with plasmids were harvested at 7 days after plasmid injection, and cells analyzed by flow cytometry using the corresponding FITC- or PE- or PE-Cy5-labeled antibodies and isotype control.

Article Snippet: The 11 kb mIL-18 DNA expression plasmid vector, pCEP4-mIL18, was constructed using a CMV early enhancer/promoter/EBNA-1-based pCEP4 plasmid vector (Invitrogen, San Diego, CA) with an ampicillin selection gene.

Techniques: Injection, Plasmid Preparation, Flow Cytometry, Labeling

Journal: BMC Cancer

Article Title: Intratumoral delivery of IL-18 naked DNA induces T-cell activation and Th1 response in a mouse hepatic cancer model

doi: 10.1186/1471-2407-7-87

Figure Lengend Snippet: IL-18 gene transfer results in regression of CT26 liver tumors . CT26 cells (1.0 × 10 5 ) were injected into a left lobe of the livers of 6-8-week-old BALB/c mice. pCEP4 or mIL-18 plasmid DNA was injected directly into the tumors on day 7. Tumor growth was measured on days 1, 4, 7, and 14 after plasmid implantation. Data are presented as the mean of tumor weights of 8 mice/group.

Article Snippet: The 11 kb mIL-18 DNA expression plasmid vector, pCEP4-mIL18, was constructed using a CMV early enhancer/promoter/EBNA-1-based pCEP4 plasmid vector (Invitrogen, San Diego, CA) with an ampicillin selection gene.

Techniques: Injection, Plasmid Preparation

Journal: BMC Cancer

Article Title: Intratumoral delivery of IL-18 naked DNA induces T-cell activation and Th1 response in a mouse hepatic cancer model

doi: 10.1186/1471-2407-7-87

Figure Lengend Snippet: Intracellular IFN-γ production by CT26-specific T cells from pmIL-18-treated mice . Splenocytes from CT26-tumor bearing mice treated with three rounds of pCEP4 or mIL-18 were pooled and incubated in medium alone or that containing CT26 lysates. Numbers within the gates in the FACS plots depict the percentage of CD69 + IFN-γ + CD4 + cells. Data are presented from experiments that were repeated at least twice.

Article Snippet: The 11 kb mIL-18 DNA expression plasmid vector, pCEP4-mIL18, was constructed using a CMV early enhancer/promoter/EBNA-1-based pCEP4 plasmid vector (Invitrogen, San Diego, CA) with an ampicillin selection gene.

Techniques: Incubation

Journal: PLoS ONE

Article Title: A Universal System for Highly Efficient Cardiac Differentiation of Human Induced Pluripotent Stem Cells That Eliminates Interline Variability

doi: 10.1371/journal.pone.0018293

Figure Lengend Snippet: ( A ) Representative immunocytochemistry of pluripotency markers POU5F1 (OCT4), NANOG, TRA-1-81, and SSEA4 in hiPSC line CBiPSC6.2 after >20 passages. ( B ) Representative hematoxylin and eosin staining of teratoma sections derived from CBiPSC6.2 after >20 passages demonstrating ectodermal, endodermal and mesodermal lineage differentiation. All CBiPSC clones in these studies formed similar teratomas. ( C ) Real-time RT-PCR studies p15 CBiPSC lines for endogenous pluripotency genes using primers that distinguish endogenous expression from transgenes (see ). ( D ) The presence of plasmid transgene sequences examined by PCR at p11 in CBiPSC6.2, CBiPSC6.11, CBiPSC6.13, CBiPSC19.11 (lanes 3–6, respectively) and negative control H9 hESC (p48) (lane 1) compared to positive control early cultures from p2 (lane 2). RT-PCR analysis of selected plasmid sequences in p11 CBiPSC6.2, CBiPSC6.11, CBiPSC6.13, CBiPSC19.11 (lanes 8–11, respectively) and negative control H9 hESC (lane 7) and p2 cultures (lane 12). ( E ) Genomic Southern blot analysis for episomal vector backbone integration in lines CBiPSC6.2, CBiPSC6.11, CBiPSC6.13, CBiPSC19.11 (p15) (lanes 2–5, respectively), H9 hESC (p55) (lane 1). Combination 6 episomal vector DNA was diluted as positive control to the equivalents of 0.4 and 4 integrations per haploid genome (0.4× and 4×). L: 1 kb plus ladder. These studies were also conducted for non-viral adult fibroblast-derived hiPSC lines iPSCWT2 and iPSCWT4 with similar results (Machairaki et al. , in preparation).

Article Snippet: EBNA-based pCEP4 vectors pEP4 EO2S EN2L ( OCT4, SOX2, NANOG, LIN28 ), pEP4 EO2S ET2K ( OCT4, SOX2, SV40LT, KLF4 ), pEP4 EO2S EM2K ( OCT4, SOX2, MYC, KLF4 ) were obtained from Addgene (Cambridge, MA, http://www.addgene.org ).

Techniques: Immunocytochemistry, Staining, Derivative Assay, Clone Assay, Quantitative RT-PCR, Expressing, Plasmid Preparation, Negative Control, Positive Control, Reverse Transcription Polymerase Chain Reaction, Southern Blot