Functional type I IFN was quantified using the reporter cell line HEK-Blue IFN-α/β according to the manufacturer’s instructions (Invivogen). Luciferase assay For ARE-Luciferase assays, experiments were performed using the calcium phosphate transfection method. For Are luciferase assays, HEK293T ...
Nrf2 reporter cell lines were prepared as previously described16. Luciferase assay Nrf2 reporter cells (5 × 105 cells/well) under G418 (100 μg/ml) were incubated with various amounts of aesculin and 3-O-β-d-glycosyl aesculin for 16 h. Total cell lysate was prepared, and ...
ciferase reporter and this event was closely correlated with the suppression of IB phosphorylation[52] . Similarly, we found that the exposure of SFN and PEITC to human pros- dimeric transcription factor that consists of the basic leu- tate cancer PC-3 cells strongly suppressed NF-B target ...
2利用H460-ARE报告基因试验细胞株为模 型发现新的Nrf2通路的抑制剂和阻遏物 利用Nrf2诱导机制,我们设计了专门受 Nrf2调控的ARE并构建r ARE一荧光素酶报告 基冈载体,率先以H460细胞株为模型建立了 H460一ARE报告基凶试验(reporter gene assay) 细胞株(专利申报中).实验表明,通过特异的 小十扰RNA降低Nrf2的表达...
We first confirmed that Brusatol could significantly inhibit NRF2 ARE reporter activity in less than 100 nM range and subsequently downregulated NRF2 target genes like HO-1 and GR. Our data shows that EAC cancer cells are sensitive to Brusatol with IC 50 within 100 nM (using Colony formation ...
BID Rabbit 2002 Cell Signalling Phospho-ERK p44/p22 Rabbit 4379 Cell Signalling Heme oxygenase-1 (HO-1) Rabbit Sc-10789 Santa Cruz Alexa Fluor 488 conjugated secondary antibody Rabbit ab150077 Abcam Alexa Fluor 568 conjugated secondary antibody Rabbit ab175471 Abcam 2.5. Luciferase Reporter As...
cell membrane damage. We predicted the binding site of Nrf2 to G6PDH by bioinformatics, which was highly conserved in various organisms. Subsequently, we confirmed this binding site by conducting the dual-luciferase reporter assay and the ChIP assay. We concluded that O-GlcNAcylated Nrf2 promoted...
vector harboring USP16-WT or USP16 with a mutated promoter region (Mutant) and Nrf2. After 36 h, a luciferase assay was performed. Unexpectedly, overexpression of Nrf2 even upregulated the luciferase reporter expression from USP16-WT, but not from USP16 Mutant (Fig.8F). However, in H/...
For this we transfected rodent F98 and human U87 glioma cells with six different expressions constructs: reporter plasmid vector (Ctrl vector), Nrf2 wild-type overexpressing vector (Nrf2 OE), dominant negative Nrf2 expression vector (Nrf2 DN) and Nrf2 siRNA knockdown vector (Nrf2 KD). Moreover,...
Ferroptosis is a distinctive process of cellular demise that is linked to amino acid metabolism, lipid oxidation, and iron oxidation. The ferroptosis cascade genes, which are closely associated with the onset of lung diseases, are among the regulatory ta