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All (9)
Promega
Addgene
Thermo Fisher
ERBA Diagnostics
MilliporeSigma
Nippon Gene Co
OriGene
SERVA
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About 100 products / 3,398 snippets / 99 images

92 / 100
Bioz Stars
pGL2 (basic) 3xARE Lux (Plasmid #14934)
Addgene  
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Catalog#: 14934
Techniques
Luciferase, Plasmid Preparation, Transfection, Software, Activity Assay (see more)
Article Snippets
".. or pU6-m3 control plasmid (Dolph Hatfield, NIH), pGL2 (basic) 3xARE Lux (Joan Massague, Memorial Sloan Kettering, Addgene # 14934), and respiratory syncytial virus (RSV)-β-galactosidase (Dr. Y. .."  (see more)
92 / 100
Bioz Stars
pGL2 basic (Plasmid #29668)
Addgene  
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Catalog#: 29668
Techniques
Plasmid Preparation, Clone Assay, Luciferase, Mutagenesis, Construct (see more)
Article Snippets
"PGL2 basic vector were obtained from Addgene."  (see more)
95+  more images
AML1/ETO transactivates the BCL-2 promoter. U937 cells were cotransfected with <t>pGL2-basic</t> or pGL2-BCL-2 reporter gene constructs and with pCMV-AML1A, pCMV-AML1B or pCMV-AML1/ETO expression vectors (together with pGHX5 to normalize transfection efficiency). Promoter activity was measured as the ratio of the luciferase activity and the concentration of growth hormone in the conditioned media. The promoter activity in the presence of the empty CMV5 plasmid was defined as 1 and the promoter activity in the presence of expression vectors for AML1A, AML1B, and AML1/ETO was defined relative to that value. The results shown are the mean of four different experiments (SEs are shown when measurable).
CONST
EXP
TNSF
ACT-A
LUCIF
C19MC miRNAs increase OCT4 expression and decrease EMT gene expression. ( a – d ) Cumulative distribution of differentially expressed mRNAs targeted by miR-512, miR-520, miR-1323 and miR-515 in HEK293 cells transfected with 759-sgRNA/SAM (759) compared to GFP transfected cells. ( e , f ) Venn diagram ( e ) and Hallmark gene set analysis ( f ) of the 181 common downregulated mRNAs in HEK293 cells transfected with 759-sgRNA/SAM or 620-sgRNA/SAM (620) compared to GFP control and predicted C19MC miRNAs target genes. ( g – i ) qRT-PCR analysis of the indicated genes normalized to GAPDH in HEK293 cells ( g , h ) or in HTR8/SVneo cells ( i ) transfected with the indicated gRNA. ( j ) Luciferase reporter assay of HEK293 cells transfected with 759-sgRNA/SAM and <t>PGL2</t> plasmid containing CDH2 3′ UTR normalized to GFP control. Graphs represent means ± SEM of at least 3 independent experiments containing 3 replicates each. * p < 0.05 vs . GFP transfected control cells. Multiple t-tests with Holm-Sidak correction ( g , I , j ); Two-way ANOVA with Dunnett’s post-hoc ( h ).
EXP
GENE-E
TNSF
CON
qRT-PCR
A) Schematic representation of three reporter vectors containing fragments of the B-MYB promoter: <t>B-MYBp-pGL2</t> (from position −102 to −709, relative to the transcription start site); 5’-Δ360 (from position −102 to position −360); 5’-Δ240 (from position −102 to −240). B) Quantifications of luciferase assays showing the activity of the B-MYB promoter segments in the presence or absence of exogenously expressed MYCN. Error bars indicate standard deviations and the asterisk indicates statistically significant differences [* = p < 0.001] with respect to the levels of B-MYBp-pGL2 promoter activity, which was arbitrarily set to 1. C) Real time PCR analysis of B-MYB expression in 293 cells transiently transfected with a MYCN expression vector. The expression levels of four independent MYCN transfectants relative to cells transfected with empty vector are shown. Error bars indicate standard deviations obtained from triplicate assays. D) Western blot analysis showing enhanced expression of B-MYB caused by activation of MYCN in SKANAS::MYCN(ER) cells. (+ or −) 4-OHT indicates that the cells were cultured in the presence or absence of 4-hydroxytamoxifen, respectively.
LUCIF
ACT-A
qPCR
EXP
TNSF
Mapping of the 5′ ends of the gene 50 transcripts by S1 nuclease protection. Analysis of transcription initiation from the putative gene 50 promoter in transiently transfected NIH 3T12 or RAW cells. The bp 66242 to 66652 gene 50 promoter-driven luciferase reporter construct, or the parent <t>pGL2</t> Basic reporter construct (Promega), was transfected into NIH 3T12 and RAW cells by the lipid-based transfection reagent SuperFect (Qiagen) as described in the legend to Fig. ​Fig.4.4. RNA was prepared (3) from cells harvested 48 h posttransfection, and 40 μg of total RNA was hybridized with 4 ng of the indicated single-stranded oligonucleotide S1 nuclease probes, which had been 32P labeled at the 5′ end with T4 polynucleotide kinase (Boehringer Mannheim, Indianapolis, Ind.) (24, 46). The sequences of the probes used were as follows: S11, 665315′-GTTTCAATTCTCATGGTCACATCTGACAGAGAAAAGGAACAGTATGAGAAATTTATGAAC-3′66472; S12, 665015′-GAAAAGGAACAGTATGAGAAATTTATGAACATACTTAAGAATCTTTCAAATTGTACTGAT-3′66442 (see Fig. ​Fig.44 for locations of the S11 and S12 probes). Following hybridization overnight at 42°C, the reaction mixture was digested with 300 U of S1 nuclease (Promega) as previously described (24, 46). The protected fragments were recovered and fractionated on a 10% denaturing acrylamide gel. Chemical cleavages of the 32P-labeled oligonucleotide probes (G + A rxn) were used as size markers in the indicated lanes. The right panel shows an analysis of transcription initiation from the gene 50 promoter in γHV68-infected NIH 3T12 cells. NIH 3T12 cells were either mock infected or infected at a multiplicity of infection of 7 in the presence of cycloheximide (final concentration, 40 μM) and anisomycin (final concentration, 10 μM) (+ CHX). Total cellular RNA was prepared (3) from cells harvested 8 h postinfection, followed by isolation of polyadenylated RNA with the PolyA Spin mRNA Isolation kit (New England Biolabs, Beverly, Mass.). Ten micrograms of poly(A) RNA isolated from mock-infected or γHV68-infected cells was hybridized with 32P-labeled S12 probe, followed by digestion with S1 nuclease as described above. In parallel, total RNA isolated from NIH 3T12 and RAW cells transfected with the bp 66242 to 66652 open reading frame 50 luciferase reporter construct was also hybridized with the S12 probe, as described above. Protected fragments were fractionated on a denaturing 10% acrylamide gel.
TNSF
LUCIF
CONST
LBLNG
HYBRID
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92 / 100
Bioz Stars
PGL2-basic
Promega  
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Techniques
 
Article Snippets
"Subsequently the SacI/XhoI fragment was subcloned into SacI/XhoI-digested pGL2-basic (Promega)."  (see more)
91 / 100
Bioz Stars
PGL2-Basic
Promega  
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Techniques
 
Article Snippets
"After overnight culture, cells were transfected with 0.05 μg pGL2-Control and 0.55 μg pGL2-Basic (Promega) using Lipofectamine 2000."  (see more)
91 / 100
Bioz Stars
PGL2-Basic
Promega  
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Techniques
 
Article Snippets
"The E-cad-luc, containing the promoter region of E-cadherin in pGL2-Basic (Promega, Cat No. E1641) was described before [ ]."  (see more)
80 / 100
Bioz Stars
PGL2-Basic
Promega  
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Techniques
 
Article Snippets
"A 1675 boIFNT1 control region ( 1675 to 66 bp) (32) was subcloned into the XhoI site of the luciferase (luc) reporter plasmid, pGL2-Basic ( 1675 luc; Promega, Madison, WI)."  (see more)
85 / 100
Bioz Stars
PGL2-Basic
Thermo Fisher Scientific / Life Technologies  
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Techniques
 
Article Snippets
".. separately transfected into the cells over 24 hours using Lipofectin reagent as indicated by the manufacturer (Life Technologies, Rockville, MD)."  (see more)
85 / 100
Bioz Stars
PGL2-Basic
Addgene  
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Techniques
 
Article Snippets
"The 1585/ 88 ID1-luc construct in pGL2-Basic (49) was purchased from Addgene (plasmid 16048)."  (see more)
85 / 100
Bioz Stars
PGL2-Basic
Addgene  
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Techniques
 
Article Snippets
".. promoter fragment (from -278) subcloned into the firefly luciferase reporter vector, pGL2-Basic (TF -278), was obtained from Addgene (Cambridge, MA, USA) 21 , 22 ."  (see more)
85 / 100
Bioz Stars
Promoterless pGL2 Basic
Promega  
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Techniques
 
Article Snippets
"As a negative control, the promoterless pGL2 Basic (Promega) was utilized."  (see more)
85 / 100
Bioz Stars
PGL2 basic reporter
Promega  
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Techniques
 
Article Snippets
"For assays of the luciferase reporter gene, four repeats of the C-1 site were subcloned into the KpnI-XhoI site of pGL2 basic reporter (Promega) with a b-actin promoter."  (see more)
85 / 100
Bioz Stars
PGL2 basic vectors
Promega  
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Techniques
 
Article Snippets
"These fragments were inserted into pGL2 basic vectors (Promega, Madison, WI), by using a Ligation kit version II® (Takara)."  (see more)