Journal: Oncotarget

Article Title: A TET2 rs3733609 C/T genotype is associated with predisposition to the myeloproliferative neoplasms harboring JAK2 V617F and confers a proliferative potential on erythroid lineages

doi: 10.18632/oncotarget.7072

Figure Lengend Snippet: ( A ) A schematic diagram of the reporter constructs containing the sequences of TET2 exon 9. The mutant construct containing identified mutation site (position +96) in the TET2 exon 9 is shown. ( B ) Results are shown as fold change of luciferase activity corresponding to pGL3-promoter vector, the wild vector displayed the stronger luciferase activity than the pGL3-promoter-vector ( P = 0.014); the luciferase activity of mutant vector was dramatically reduced to 0.38 fold in transfected HEL cells than that of the wild-type vector ( P = 0.009). Values are means ± SD of triplet data from 3 different experiments.

Article Snippet: To further testify whether rs3733609 C/T genotype is able to affect the transcriptional activity of the TET2 gene, a corresponding reporter gene plasmid was constructed as follows: 1). pGL3-promoter vector (empty vector, Promega, USA), containing the SV40 promoter of driving the firefly luciferase gene; 2). pGL3-pro-TET2-exon9-wt, containing the luciferase reporter gene and carrying 138 bp of TET2 exon 9 (rs3733609 T/T genotype); 3). pGL3-pro-TET2-exon9-Mut96 (rs3733609 C/T genotype), containing the TET2 exon 9 c.138 + 96T > C variation.

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

Journal: Oncogenesis

Article Title: Overexpression of miR-26a-2 in human liposarcoma is correlated with poor patient survival

doi: 10.1038/oncsis.2013.10

Figure Lengend Snippet: Effect of miR-26a-2 on adipocyte differentiation and apoptosis in LPS cells. For panels ( a ), ( b ) and ( d ), LPS cells were transduced with either pMSCV-miR-26a-2 expression vector (26A2) or empty vector control. Stable clones were selected with puromycin. For panels ( c ) and ( e ), LPS cells were transfected with either scrambled control oligos or anti-miR-26a-2 oligos. Cells were harvested 24 h after transfection. ( a ) mRNA expression level of adipocyte differentiation marker genes in miR-26a-2 overexpression clones. Dashed line indicates the normalized expression level of each gene compared with the empty vector clones. ( b ) Western blots showing protein expression level of C/EBPβ, C/EBPα and PPARγ in miR-26a-2 overexpression clones. GAPDH was used as a loading control. ( c ) Protein expression level of C/EBPβ, C/EBPα and PPARγ in LPS cells transfected with anti-miR-26a-2 oligos. ( d ) Apoptosis assay results of miR-26a-2 overexpression clones after 72 h of exposure to nocodazole. ( e ) Apoptosis assay results of cells transfected with anti-miR-26a-2 after 72 h exposure to nocodazole. Experiments were repeated three times to ensure accuracy. Data represent average±s.d. (error bars). Asterisks (* and **) indicate P -values <0.05 and 0.001 by t -test, respectively.

Article Snippet: Cotransfection of pGL3-Promoter-RCBTB1 3′-UTR vector and pRL-TK renilla luciferase vector, along with either miR-26a-2 expression vector or empty vector control, was performed using Bio-T transfection reagent according to the manufacturer's protocol (BioLand LLC, Paramount, CA).

Techniques: Transduction, Expressing, Plasmid Preparation, Control, Clone Assay, Transfection, Marker, Over Expression, Western Blot, Apoptosis Assay