His lab crossed the K - ras model to a ubiquitous promoter driving conditional
luciferase expression only in cells that Cre has entered.
All three compounds were able to induce
luciferase expression, demonstrating their ability to overcome Bach1 repression of HMOX E2 - dependent transcription (Figure 6C).
Nonetheless, CDDO - Me and HPP - 1014 still were able to activate HMOX1 E2 - dependent
luciferase expression in the presence of mutant Bach1 proteins, indicating that the CP motifs in Bach1 are not critical for efficient derepression of Bach1 by an electrophile (Figure 6D).
Expression of HMOX1 E2 - dependent
luciferase expression was determined in HepG2 cells co-transfected with an HMOX1 E2 - dependent reporter plasmid and a plasmid expressing FLAG - tagged wildtype Bach1 (Figure 6A).
Luciferase expression was markedly lower in cells expressing Bach1, indicating effective repression of HMOX1 E2 - dependent transcription by Bach1 (Figure 6B).
In these assays, both wild - type Bach1 and FLAG - hBach1 - AP4 - 7 efficiently repressed basal levels of
luciferase expression.
In contrast, the ability of both CoPP and HPP - 4382 to induce HMOX1 E2 ARE - dependent
luciferase expression in the presence of the mutant Bach1 protein was sharply inhibited.
When compared to unfractionated hu - PBMCs, the CD8 + - depleted hu - PBMC recipients show similar reductions in
luciferase expression following IL - 15 superagonist treatment.
When spleens from infected mice were examined for sustained control of infection at later time points, however,
luciferase expression had rebounded.
(11) To more directly test for defects in Wnt signaling, CAST / EiJ and C57BL / 6J WT and mTERT — / — embryonic fibroblasts were transfected with a luciferase reporter plasmid with a Wnt3a ligand;
luciferase expression was indistinguishable between WT and mTERT — / — cells.
Cells were transfected with 320 ng pGL3
luciferase expression construct containing the 3 ′ UTR of human APC, 40 ng pGL4.74 hRLuc / TK Renilla luciferase vector (Promega, Fitchburg, WI), and 25 nM hsa - miR - 142 precursor or negative control precursor (Ambion).
Not exact matches
Ten nanograms of reporter plasmid and 200 ng of miR7
expression plasmid were cotransfected with 1 ng of phRG - TK Renilla
luciferase internal control plasmid (Promega) into 293TN cells using Lipofectamine 2000 reagent (Invitrogen).
Cells were transfected with the human ID1 promoter -LRB--- 985 / +94)
luciferase reporter construct (31) together with the wild - type or mutant ACVR1
expression construct.
To examine whether DDX3 transactivates the p21waf1 / cip1 promoter activity through Sp1 site - interacting proteins, the Sp1 or Sp3
expression plasmid together with the DDX3
expression construct and the p21waf1 / cip1 promoter — driven
luciferase reporter were introduced into HuH - 7 cells.
p21waf1 / cip1 promoter — driven
luciferase reporter (p21 - Luc; 0.05 - 0.5 μg) and increasing amount (0.1 - 2 μg) of FLAG - DDX3
expression construct were cotransfected into various cell lines as indicated.
As shown in Fig. 2A, exogenous
expression of DDX3 in various cell lines led to a 2 - to 4-fold up - regulation of p21waf1 / cip1 promoter — driven
luciferase activity.
To this end, increasing amounts of
expression constructs of DDX3 / DQAD, DDX3 / AAA and wild - type DDX3 were cotransfected with p21waf1 / cip1 promoter — driven
luciferase reporter.
The cells were cotransfected with the reporter gene constructs and related
expression plasmids as indicated by using SuperFect (Qiagen) followed by transcriptional activity assay using Dual -
Luciferase Assay System (Promega) according to the manufacturer's instruction.
Second, in IRF - 9 — deficient U2A cells, IFNα - induced
luciferase activity was detected only when enforcing IRF - 9
expression in these cells (Fig. 1D).
In agreement with this, both wt STAT2 and mutant STAT2 - Y690F displayed a similar effect on the
luciferase reporter gene
expression in U3A cells without IFNα.
Gene
expression Reporter genes, such as
luciferase or GFP, can also be assessed in microplate readers, enabling in vitro and in vivo determination of gene
expression for studies using markers of genetic alteration.
The AG haplotype showed significantly greater
expression of
luciferase than the GT haplotype.
The minor allele of rs25532 significantly decreased
luciferase reporter gene
expression levels by 15 — 80 %, depending on 5 - HTTLPR allele background and cell type.