Journal: Scientific Reports

Article Title: Piperlongumine inhibits lung tumor growth via inhibition of nuclear factor kappa B signaling pathway

doi: 10.1038/srep26357

Figure Lengend Snippet: ( a,b ) NSCLC cells were treated with PL (0–15 μM) for 2 h, and then were lysed with A buffer and C buffer. Nuclear extracts were incubated in binding reactions of 3 2 p-end-labeled oligo nucleotide containing the NF-κB sequence. The present EMSA results are representative for three experiments. ( c,d ) NSCLC cells were tranfected with pNF-κB-Luc plasmid for 24 h. The transfected cells were treated with PL (0–15 μM) for another 24 h. Luciferase activity was measured by using the luciferase assay kit. The present results are representative for three experiments. ( c , d ) Cytosol proteins were used to determine the expression of IκBα, p-IκBα and β-actin (internal control) in NSCLC cells. Nuclear proteins were used to determine the expression of p50, p65 and Histone H1 (internal control) in NSCLC cells. For the cropped images, samples were run in the same gels under same experimental conditions and processed in parallel. Each band is representative for three experiments. Full-length gels are presented in .

Article Snippet: The DNA binding activity of NF-κB was determined using an electrophoretic mobility shift assay (EMSA) performed as according to the manufacturer’s recommendations (Promega, Madison, USA).

Techniques: Incubation, Binding Assay, Labeling, Sequencing, Plasmid Preparation, Transfection, Luciferase, Activity Assay, Expressing, Control

Journal: Scientific Reports

Article Title: Piperlongumine inhibits lung tumor growth via inhibition of nuclear factor kappa B signaling pathway

doi: 10.1038/srep26357

Figure Lengend Snippet: ( a ) Cells were transfected with p50 mutant plasmid (C62S) for 24 h, then were treated with PL (10 μM) for another 24 h. Cell viability was determined by MTT assay. Data was expressed as the mean ± S.D. of three experiments. * p < 0.05 indicates significantly different from control cells. # p < 0.05 indicates significantly different from PL-treated cells. ( b ) Effect of p50 mutant plasmid (C62S) on the expression of DRs was determined by using Western blotting with antibodies against Fas, DR4 and β-actin (internal control). Each band is representative for three experiments. ( c ) Cells were transfected with Fas siRNA or DR4 siRNA for 24 h, and then were treated with PL (10 μM) for another 24 h. Cell viability was determined by MTT assay. Data was expressed as the mean ± S.D. of three experiments. * p < 0.05 indicates significantly different from control cells. # p < 0.05 indicates significantly different from PL-treated cells. ( d ) Effect of Fas siRNA or DR4 siRNA transfection on DNA binding activity of NF-κB was determined by EMSA as described in the materials and methods. For the cropped images, samples were run in the same gels under same experimental conditions and processed in parallel. Full-length gels are presented in .

Article Snippet: The DNA binding activity of NF-κB was determined using an electrophoretic mobility shift assay (EMSA) performed as according to the manufacturer’s recommendations (Promega, Madison, USA).

Techniques: Transfection, Mutagenesis, Plasmid Preparation, MTT Assay, Control, Expressing, Western Blot, Binding Assay, Activity Assay

Journal: Scientific Reports

Article Title: Piperlongumine inhibits lung tumor growth via inhibition of nuclear factor kappa B signaling pathway

doi: 10.1038/srep26357

Figure Lengend Snippet: ( a ) Expression of DR was determined by Western blotting with antibodies against Fas, DR4 and β-actin (internal control). ( b ) Expression of apoptosis regulatory proteins was determined by Western blotting with antibodies against cleaved capase-3, cleaved caspase-8, Bax, Bcl-2 and β-actin (internal control). ( c ) Lung tumors were lysed with A buffer and C buffer. Nuclear extracts were incubated in binding reactions of 3 2 p-end-labeled oligo nucleotide containing the NF-κB sequence. The present EMSA results are representative for three experiments. ( d ) Cytosol proteins were used to determine the expression of IκBα, p-IκBα and β-actin (internal control). Nuclear proteins were used to determine the expression of p50, p65 and Histone H1 (internal control) in lung tumor tissues. For the cropped images, samples were run in the same gels under same experimental conditions and processed in parallel. Each band is representative for three experiments. Full-length gels are presented in and .

Article Snippet: The DNA binding activity of NF-κB was determined using an electrophoretic mobility shift assay (EMSA) performed as according to the manufacturer’s recommendations (Promega, Madison, USA).

Techniques: Expressing, Western Blot, Control, Incubation, Binding Assay, Labeling, Sequencing