plasmid dnas encoding human genes  (GenScript corporation)

Journal: Cancers

Article Title: Epigenetic Silencing of miR-9 Promotes Migration and Invasion by EZH2 in Glioblastoma Cells

doi: 10.3390/cancers12071781

Figure Lengend Snippet: High expression of EZH2 and CXCR4 is associated with the poor prognosis of glioblastoma (GBM) patients. ( A ) EZH2 and CXCR4 mRNA levels are significantly higher in GBM tumor tissues than normal tissues (tumor samples, n = 163; normal samples, n = 207). * p < 0.05: significantly different from normal tissues. ( B ) Both high EZH2 or CXCR4 levels correspond with the reduced disease-free survival of GBM patients based on data obtained from Gene Expression Profiling Interactive Analysis (GEPIA).

Article Snippet: The DNA plasmids encoding CXCR4 were from Sino Biological (cat. HG11325-CF).

Techniques: Expressing

Journal: Cancers

Article Title: Epigenetic Silencing of miR-9 Promotes Migration and Invasion by EZH2 in Glioblastoma Cells

doi: 10.3390/cancers12071781

Figure Lengend Snippet: Upregulation of CXCR4 expression by EZH2. ( A ) Differential expression of EZH2 and CXCR4 mRNA in U87-MG, GBM8401, and DBTRG-05MG cells. The expression of EZH2 and CXCR4 was determined using qRT-PCR. ( B ) Western blot analysis of EZH2, CXCR4, and actin (loading control) in whole-cell lysates. ( C ) The effect of shEZH2 or Myc-EZH2 on mRNA expression in GBM8401, DBTRG-05MG, and U87-MG cells was evaluated using qRT-PCR. CXCR4 mRNA expression is shown relative to control. ( D ) Western blot analysis of EZH2 and CXCR4 in EZH2 knockdown/overexpression (OE) GBM cells. (mean ± SD, n = 3). * p < 0.05. The raw data of Western blots is shown in .

Article Snippet: The DNA plasmids encoding CXCR4 were from Sino Biological (cat. HG11325-CF).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Over Expression

Journal: Cancers

Article Title: Epigenetic Silencing of miR-9 Promotes Migration and Invasion by EZH2 in Glioblastoma Cells

doi: 10.3390/cancers12071781

Figure Lengend Snippet: CXCR4 is a target of miR-9. ( A ) The intersection of the analyses using the three algorithms (miRWalk, Pictar, and TargetScan) is shown for the miRNAs that were predicted to target CXCR4 and those that were differentially regulated. ( B ) Binding site of miR-9 predicted in the CXCR4 3′ UTR by TargetScan. ( C ) Differential expression of miR-9 in U87-MG, GBM8401, and DBTRG-05MG cells. The expression of miR-9 was determined using qRT-PCR. ( D ) The effect of the miR-9 mimic (10 nM) in the indicated cells was evaluated using qRT-PCR. ( E ) The relative CXCR4 mRNA levels in GBM8401 and DBTRG-05MG cells 48 h after transfection of the miR-9 mimic (10 nM) or miR-Ctrl ( F ) The protein levels of CXCR4 were measured by Western blotting. β-actin was used as an endogenous control. ( G ) The effect of the miR-9 inhibitor (10 nM) on the miR-9 level in U87-MG cells was evaluated using qRT-PCR. Cells were transfected with miR-9 inhibitor or control, and CXCR4 mRNA ( H ) and protein ( I ) were determined 48 h after transfection. Mean ± SD, n = 3. * p < 0.05; ** p < 0.01. The raw data of Western blots is shown in .

Article Snippet: The DNA plasmids encoding CXCR4 were from Sino Biological (cat. HG11325-CF).

Techniques: Binding Assay, Expressing, Quantitative RT-PCR, Transfection, Western Blot

Journal: Cancers

Article Title: Epigenetic Silencing of miR-9 Promotes Migration and Invasion by EZH2 in Glioblastoma Cells

doi: 10.3390/cancers12071781

Figure Lengend Snippet: The effects of EZH2 and CXCR4 knockdown or overexpression on GBM cell migration. ( A ) Transwell migration assays were used to evaluate the effects of shEZH2-2 or Myc-EZH2 overexpression on the migration of GBM cells. Representative photographs of transwell assays. The bound crystal violet staining was released with 33% glacial acetic acid and quantified by absorbance measurements (OD570) ( B ) Western blot analysis of CXCR4 protein in CXCR4 knockdown cells. ( C ) Transwell migration assays and quantification of these assays were used to assess the effects of shCXCR4-1 and shLuc (as a control) on GBM cell migration. ( D ) Western blot analysis of CXCR4 protein in Flag-CXCR4 overexpression (OE) U87-MG cells. ( E ) Transwell migration assays and quantification of these assays were used to assess the effects of Flag-CXCR4 on U87-MG migration. Mean ± SD, n = 3. * p < 0.05; ** p < 0.01. The raw data of Western blots is shown in .

Article Snippet: The DNA plasmids encoding CXCR4 were from Sino Biological (cat. HG11325-CF).

Techniques: Over Expression, Migration, Staining, Western Blot

Journal: Cancers

Article Title: Epigenetic Silencing of miR-9 Promotes Migration and Invasion by EZH2 in Glioblastoma Cells

doi: 10.3390/cancers12071781

Figure Lengend Snippet: Proposed signaling for EZH2/miR-9/CXCR4 modulation of GBM cell migration and invasion. EZH2 modulates the expression of miR-9, which directly targets the oncogenic signaling of CXCR4 in GBM cell lines.

Article Snippet: The DNA plasmids encoding CXCR4 were from Sino Biological (cat. HG11325-CF).

Techniques: Migration, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Chitosan Particles Complexed with CA5-HIF-1α Plasmids Increase Angiogenesis and Improve Wound Healing

doi: 10.3390/ijms241814095

Figure Lengend Snippet: Luciferase expression over time when using chitosan particles for plasmid transfection. Verification of transfection efficiency of luciferase plasmids (Luc) using chitosan particles (CPs) in an in vivo reporter assay wound model. Statistical significance was calculated using two-way ANOVA with Holm–Sidak’s multiple comparison test (* p < 0.05) ( n = 3).

Article Snippet: Plasmid DNA encoding the human CA5-HIF-1α gene and firefly luciferase gene was provided by Nature Technology Corporation (NTC) (Lincoln, NE, USA). pCMV-GFP was a gift from Connie Cepko (Addgene plasmid #11153; http://n2t.net/addgene:11153 ; RRID:Addgene_11153 (accessed on 12 July 2021).

Techniques: Luciferase, Expressing, Plasmid Preparation, Transfection, In Vivo, Reporter Assay, Comparison