Journal: iScience

Article Title: DNA methyltransferases are complementary in maintaining DNA methylation in embryonic stem cells

doi: 10.1016/j.isci.2022.105003

Figure Lengend Snippet:

Article Snippet: AxyPrep Endo-Free Plasmid Midiprep Kit , Axygen of Corning , Cat. # 09318KA2.

Techniques: Virus, Recombinant, Gel Extraction, Plasmid Preparation, Methylation, Software

Journal: PLoS ONE

Article Title: Leptin Is an Anti-Apoptotic Effector in Placental Cells Involving p53 Downregulation

doi: 10.1371/journal.pone.0099187

Figure Lengend Snippet: A) Swan-71 cells (1×10 6 cells) were plated in DMEM-F12 media in the absence of serum and incubated during 72 h with different doses of leptin. DMEM-F12 10% FBS was used as a control. Cell extracts were prepared as indicated in . Proteins were separated on SDS-PAGE gels and BCL-2 and BAX expression was determined by Western blot analysis. Molecular weights were estimated using standard protein markers. Molecular mass (kDa) is indicated at the right of the blot. Loading controls were performed by immunoblotting the same membranes with anti-α-tubulin. Bands densitometry is shown in lower panels, results are expressed as mean ± SD for three independent experiments. B) Leptin increased BCL-2/BAX relationship. C) BeWo cells were transiently transfected with a plasmid containing a section of BAX promoter (pBax-Luc). After transfection, cells were incubated for 72 h in DMEM-F12 and treated with increasing leptin doses. Cell extracts were prepared as indicated in and Luciferase activity was normalized to β-galactosidase activity. Activity obtained in the absence of leptin and pBax-Luc was set as control. Statistical analyzes were performed by ANOVA. Asterisks indicate significant differences from the control according to Bonferroni's multiple comparison post hoc test, relative to FBS 10% (#) or FBS 0% (*). ## p<0.01, * p<0.05, *** p<0.001.

Article Snippet: To perform transient transfection assays, plasmids were purified using the Midipreps Wizard kit (Promega Co.), and DNA concentration was estimated spectrophotometrically.

Techniques: Incubation, Control, SDS Page, Expressing, Western Blot, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Comparison

Journal: Cytotechnology

Article Title: Production of lentiviral vectors in suspension cells using low proportion of supercoiled circular plasmid DNA

doi: 10.1007/s10616-020-00433-4

Figure Lengend Snippet: Agarose gel electrophoresis of SC, OC and linear forms of four plasmids. Lane 1, native plasmid DNA of VSVG; lane 2, 7-day treatment at 50 °C for plasmid DNA of VSVG; lane 3, a mixture of native VSVG plasmid DNA and heat-treated VSVG plasmid DNA at ratio of 1:1; lane 4, native plasmid DNA of MDK; lane 5, 7-day treatment at 50 °C for plasmid DNA of MDK; lane 6, a mixture of native MDK plasmid DNA and heat-treated MDK plasmid DNA at ratio of 1:1; lane 7, native plasmid DNA of RSK; lane 8, 7-day treatment at 50 °C for plasmid DNA of RSK; lane 9, a mixture of native RSK plasmid DNA and heat-treated RSK plasmid DNA at ratio of 1:1; lane 10, native plasmid DNA of pLenti6.3/V5-CD19 CAR; lane 11, 7-day treatment at 50 °C for plasmid DNA of pLenti6.3/V5-CD19 CAR; lane 12, a mixture of native pLenti6.3/V5-CD19 CAR plasmid DNA and heat-treated pLenti6.3/V5-CD19 CAR plasmid DNA at ratio of 1:1

Article Snippet: The next day, the plasmid DNA was extracted using the AxyPrep Plasmid Midiprep Kit (Axygen, Corning Incorporated, USA).

Techniques: Agarose Gel Electrophoresis, Plasmid Preparation

Journal: Cytotechnology

Article Title: Production of lentiviral vectors in suspension cells using low proportion of supercoiled circular plasmid DNA

doi: 10.1007/s10616-020-00433-4

Figure Lengend Snippet: Effect of the SC plasmid DNA proportion on the functional titer of lentiviral vectors produced by HEK293T cells. Functional titer was measured by transduction units (TU). Lentivirus samples in three groups were harvested individually. Error bars represent the standard deviation of three independent infections. *P < 0.05; **P < 0.01

Article Snippet: The next day, the plasmid DNA was extracted using the AxyPrep Plasmid Midiprep Kit (Axygen, Corning Incorporated, USA).

Techniques: Plasmid Preparation, Functional Assay, Produced, Transduction, Standard Deviation

Journal: Cytotechnology

Article Title: Production of lentiviral vectors in suspension cells using low proportion of supercoiled circular plasmid DNA

doi: 10.1007/s10616-020-00433-4

Figure Lengend Snippet: Effect of the SC plasmid DNA proportion on the functional titer of lentiviral vectors produced by HEK293SF cells. Functional titer was measured by transduction units (TU). Lentivirus samples in three groups were harvested individually. Error bars represent the standard deviation of three independent experiments. *P < 0.05; **P < 0.01; NS, no significant difference

Article Snippet: The next day, the plasmid DNA was extracted using the AxyPrep Plasmid Midiprep Kit (Axygen, Corning Incorporated, USA).

Techniques: Plasmid Preparation, Functional Assay, Produced, Transduction, Standard Deviation

Journal: Cytotechnology

Article Title: Production of lentiviral vectors in suspension cells using low proportion of supercoiled circular plasmid DNA

doi: 10.1007/s10616-020-00433-4

Figure Lengend Snippet: Agarose gel electrophoresis of SC, OC and linear forms of four plasmids after heat or enzyme treatment. Lane 1, native plasmid DNA of RSK; lane 2, 7-day treatment at 50 °C for plasmid DNA of RSK; lane 3, 10-day treatment at 50 °C for plasmid DNA of RSK; lane 4, RSK plasmid DNA was digested with Xho I; lane 5, native plasmid DNA of VSVG; lane 6, 7-day treatment at 50 °C for plasmid DNA of VSVG; lane 7, 10-day treatment at 50 °C for plasmid DNA of VSVG; lane 8, VSVG plasmid DNA was digested with Hand III; lane 9, native plasmid DNA of MDK; lane 10, 7-day treatment at 50 °C for plasmid DNA of MDK; lane 11, 10-day treatment at 50 °C for plasmid DNA of MDK; lane 12, MDK plasmid DNA was digested with Mun I; lane 13, native plasmid DNA of pLenti6.3/V5-CD19 CAR; lane 14, 7-day treatment at 50 °C for plasmid DNA of pLenti6.3/V5-CD19 CAR; lane 15, 10-day treatment at 50 °C for plasmid DNA of pLenti6.3/V5-CD19 CAR; lane 16, pLenti6.3/V5-CD19 CAR plasmid DNA digested with Sal I

Article Snippet: The next day, the plasmid DNA was extracted using the AxyPrep Plasmid Midiprep Kit (Axygen, Corning Incorporated, USA).

Techniques: Agarose Gel Electrophoresis, Plasmid Preparation