cell nuclear antigen inhibitor  (Tocris)

Journal: Antioxidants

Article Title: Piperlongumine Induces Cell Cycle Arrest via Reactive Oxygen Species Accumulation and IKKβ Suppression in Human Breast Cancer Cells

doi: 10.3390/antiox8110553

Figure Lengend Snippet: Primers used for real-time PCR.

Article Snippet: An NF-κB inhibitor (Bay 11-7082), and antibodies against cyclin D1, CDK4, CDK6, proliferating cell nuclear antigen (PCNA), NF-κB p65, IκBα, lamin B, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), goat anti-rabbit IgG-HRP, and donkey anti-goat IgG-HRP were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques:

Journal: Antioxidants

Article Title: Piperlongumine Induces Cell Cycle Arrest via Reactive Oxygen Species Accumulation and IKKβ Suppression in Human Breast Cancer Cells

doi: 10.3390/antiox8110553

Figure Lengend Snippet: Effects of piperlongumine (PL) on cell cycle-regulatory proteins in MCF-7 cells. The expression of cyclin B1, p-CDK1, CDK1, PCNA, cyclin D1, CDK6, and CDK4 at mRNA ( A ) and protein levels ( B ) was measured using real time PCR and Western blotting, respectively. GAPDH was used as an internal control gene and as a loading control. Data represent the mean ± SEM (n = 3). * Significantly different compared with the control ( p < 0.05).

Article Snippet: An NF-κB inhibitor (Bay 11-7082), and antibodies against cyclin D1, CDK4, CDK6, proliferating cell nuclear antigen (PCNA), NF-κB p65, IκBα, lamin B, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), goat anti-rabbit IgG-HRP, and donkey anti-goat IgG-HRP were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Pharmaceutical biology

Article Title: Anti-neuroinflammatory effects of cudraflavanone A isolated from the chloroform fraction of Cudrania tricuspidata root bark.

doi: 10.1080/13880209.2018.1447972

Figure Lengend Snippet: Figure 4. Effect of cudraflavanone A on LPS-induced activation of NF-jB in BV2 cells. Cells were pretreated with the indicated concentrations of cudraflavanone A for 3 h and then stimulated with LPS (1 lg/mL) for 1 h. Nuclear and cytosolic extracts were isolated and the levels of p65 and p50 in the nuclear fraction, and p-IjB-a and IjB-a in the cytosolic fraction were determined by Western blot analysis. PCNA and actin were used as internal controls. The experiment was repeated three times, and similar results were obtained.

Article Snippet: Primary antibodies such as anti-iNOS, antiCOX-2, anti-inhibitor kappa B (IrB)-a, anti-p-IrB-a, anti-p50, anti-p65, anti-actin and anti-proliferating cell nuclear antigen (PCNA) antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX).

Techniques: Activation Assay, Isolation, Western Blot

Journal: Biochimica et biophysica acta

Article Title: Transient receptor potential channel M2 contributes to neointimal hyperplasia in vascular walls.

doi: 10.1016/j.bbadis.2015.03.014

Figure Lengend Snippet: Fig. 1. Neointimal hyperplasia and ROS over-production in cuff-injured rat femoral arteries. (A) Histological analysis showing cuff-induced neointimal hyperplasia in the artery. Cross sec- tions of artery were stained with hematoxylin and eosin. The hyperplasia was expressed as the ratio of the intimal area to the medial area (I/M ratio) and displayed on the right. Data are expressed as mean ± SEM. n = 5. ** p b 0.01 versus sham. (B) Cross sections of artery stained with anti-proliferating cell nuclear antigen (PCNA) antibody. n = 5. (C) Western blot of adventitial-removed artery segments probed with anti-PCNA antibody and anti-β-tubulin antibody. Data are expressed as mean ± SEM. n = 3. ** p b 0.01 versus sham. (D) Cross sections of artery stained with dihydroethidium (DHE), a ROS-sensitive fluorescent probe. The fluorescent intensity of sham-operated artery was normalized to 100%. Data are expressed as mean ± SEM. n = 4. ** p b 0.01 versus sham. Arrowheads in A and D indicate the internal elastic lamina, which forms a boundary between arterial intima and media.

Article Snippet: Akt inhibitor IV, Akt inhibitor X, mouse monoclonal anti-proliferating cell nuclear antigen (PCNA) antibody, goat polyclonal anti-Axl antibody, rabbit polyclonal anti-β-tubulin antibody, mouse monoclonal anti-β-actin antibody, and horseradish peroxidase-conjugated donkey anti-goat IgG were obtained from Santa Cruz Biotechnology.

Techniques: Staining, Western Blot

Journal: Biochimica et biophysica acta

Article Title: Transient receptor potential channel M2 contributes to neointimal hyperplasia in vascular walls.

doi: 10.1016/j.bbadis.2015.03.014

Figure Lengend Snippet: Fig. 3. Reduced neointimal hyperplasia in cuff-injured carotid artery of Trpm2 knockout mice. (A) Cross sections of artery stained with hematoxylin and eosin. Arrowheads indicate the internal elastic lamina. The hyperplasia was expressed as I/M ratio and displayed on the right. Data are expressed as mean ± SEM. n = 4. * p b 0.05, ** p b 0.01. (B) Western blot of ad- ventitial-removed artery segments probed with anti-TRPM2 antibody and anti-β-tubulin antibody. Data are expressed as mean ± SEM. n = 3. ** p b 0.01. (C) Western blot of adventi- tial-removed artery segments probed with anti-PCNA antibody and anti-β-tubulin antibody. Data are expressed as mean ± SEM. n = 4. * p b 0.05, ** p b 0.01. WT, wild-type; Trpm2 KO, Trpm2 knockout.

Article Snippet: Akt inhibitor IV, Akt inhibitor X, mouse monoclonal anti-proliferating cell nuclear antigen (PCNA) antibody, goat polyclonal anti-Axl antibody, rabbit polyclonal anti-β-tubulin antibody, mouse monoclonal anti-β-actin antibody, and horseradish peroxidase-conjugated donkey anti-goat IgG were obtained from Santa Cruz Biotechnology.

Techniques: Knock-Out, Staining, Western Blot

Journal: Biochimica et biophysica acta

Article Title: Transient receptor potential channel M2 contributes to neointimal hyperplasia in vascular walls.

doi: 10.1016/j.bbadis.2015.03.014

Figure Lengend Snippet: Fig. 5. Role of TRPM2 in H2O2-stimulated proliferation of rodent aortic SMCs. (A) and (B) EdU incorporation assay showing inhibitory effect of TM2E3 (10 μg/ml) and Trpm2 knockout on H2O2 (10 μM)-induced proliferation of rat (A) and mouse (B) aortic SMCs. Nuclei were stained with Hoechst 33342 (blue). Cell proliferation was expressed as the percentage of EdU-pos- itive cells (red). Data are expressed as mean ± SEM. n = 7 (A) and 9 (B) independent experiments. ** p b 0.01. (C) and (D) MTT assay showing inhibitory effect of TM2E3 (10 μg/ml) and Trpm2 knockout on H2O2 (10 μM)-induced proliferation of rat (C) and mouse (D) aortic SMCs. In C, pre-immune IgG group in the absence of H2O2 was normalized to 100%. Data are expressed as mean ± SEM. n = 6 independent experiments. ** p b 0.01. In D, WT group in the absence of H2O2 was normalized to 100%. Data are expressed as mean ± SEM. n = 4 in- dependent experiments. ** p b 0.01. (E) Western blot of aortic SMCs probed with anti-PCNA antibody and anti-β-tubulin antibody. Data are expressed as mean ± SEM. n = 3. ** p b 0.01. WT, wild-type; Trpm2 KO, Trpm2 knockout.

Article Snippet: Akt inhibitor IV, Akt inhibitor X, mouse monoclonal anti-proliferating cell nuclear antigen (PCNA) antibody, goat polyclonal anti-Axl antibody, rabbit polyclonal anti-β-tubulin antibody, mouse monoclonal anti-β-actin antibody, and horseradish peroxidase-conjugated donkey anti-goat IgG were obtained from Santa Cruz Biotechnology.

Techniques: Knock-Out, Staining, MTT Assay, Western Blot

Journal: Experimental & Molecular Medicine

Article Title: Polyphenols isolated from Broussonetia kazinoki prevent cytokine-induced β-cell damage and the development of type 1 diabetes

doi: 10.1038/emm.2015.16

Figure Lengend Snippet: Prevention of cytokine-induced NF-κB activation in RINm5F cells. ( a ) RINm5F cells were pretreated with the indicated concentrations of kazinol C or isokazinol D for 3 h, and followed by IL-1β and IFN-γ treatment. Following 24 h of incubation, the concentration of nitrite and the expression of iNOS mRNA and protein were determined. Each value is the mean±s.e.m. of three independent experiments ( n =9). ** P <0.01 versus vehicle-treated control; ## P <0.01 versus cytokine-treated cells. ( b ) After 30 min, NF-κB DNA binding was analyzed by electrophoretic mobility shift assay, and the nuclear translocation of the p65 and p50 subunits and the level of cytoplasmic IκBα were determined by western blotting. β-actin and proliferating cell nuclear antigen (PCNA) were used as loading controls for cytosolic and nuclear proteins, respectively.

Article Snippet: Membranes were probed with primary antibodies (1 μg ml −1 ) against p50, p65, inhibitor of κBα (IκBα), iNOS, proliferating cell nuclear antigen and β-actin (all from Santa Cruz Biotechnology, Dallas, TX, USA), and cleaved caspase-3, Bcl-2 and Bax (Cell Signaling Technology, Beverly, MA, USA).

Techniques: Activation Assay, Incubation, Concentration Assay, Expressing, Control, Binding Assay, Electrophoretic Mobility Shift Assay, Translocation Assay, Western Blot

Journal: Experimental & Molecular Medicine

Article Title: Polyphenols isolated from Broussonetia kazinoki prevent cytokine-induced β-cell damage and the development of type 1 diabetes

doi: 10.1038/emm.2015.16

Figure Lengend Snippet: Prevention of cytokine-induced NF-κB activation in isolated islets. Mouse islets were treated with IL-1β (1 U ml −1 ) and IFN-γ (100 U ml −1 ), either with or without a 3-h pretreatment with the indicated concentration of kazinol C or isokazinol D. ( a ) The concentration of nitrite and the expression of iNOS mRNA and protein were determined after 24 h. ( b ) The extents of NF-κB DNA binding, IκBα degradation and nuclear translocation of p65 and p50 were determined 1 h later. The results of triplicate samples are expressed as means±s.e.m. ( n =9). ** P <0.01 versus vehicle-treated control; ## P <0.01 versus cytokine-treated islets.

Article Snippet: Membranes were probed with primary antibodies (1 μg ml −1 ) against p50, p65, inhibitor of κBα (IκBα), iNOS, proliferating cell nuclear antigen and β-actin (all from Santa Cruz Biotechnology, Dallas, TX, USA), and cleaved caspase-3, Bcl-2 and Bax (Cell Signaling Technology, Beverly, MA, USA).

Techniques: Activation Assay, Isolation, Concentration Assay, Expressing, Binding Assay, Translocation Assay, Control