rabbit anti phospho iκb beta antibody  (Bioss)

Journal: International Journal of Biological Sciences

Article Title: CXCL14 promotes metastasis of non-small cell lung cancer through ACKR2-depended signaling pathway

doi: 10.7150/ijbs.79438

Figure Lengend Snippet: Schematic mechanism of CXCL14 promotes tumor metastasis in lung cancer. The ligand CXCL14 stimulates the atypical chemokine receptor 2 (ACKR2) to activate PLCβ3, PKCα, and c-Src. The phosphorylation of phospholipase Cβ3 (PLCβ3), protein kinase Cα (PKCα), and proto-oncogene c-Src (c-Src) signaling pathways activates IκB kinase α (IKKα) and NF-κB inhibitor α (IκBα) to release nuclear factor-κB (NF-κB). The nuclear translocation of NF-κB promotes the expression of EMT protein and cell migration that leads to tumor metastasis.

Article Snippet: Primary antibodies specific for phospho-phospholipase Cβ3 (PLCβ3; Ser537, #2481), PLCβ3 (#14247), phospho-protein kinase Cα/βII (PKCα/βII; Thr638/641, #9375), PKCα (#2056), phospho-proto-oncogene c-Src (c-Src; Ser17, #5473), c-Src (#2109), phosphor-inhibitor of NF-κB α (IκBα; Ser32/36, #9246), phosphor-IκB kinase α/β (IKKα/β; Ser176/180, #2697), phospho-NF-κB p65 (Ser536, #3033), NF-κB p65 (#8242), and ZO-1 (#8193) were purchased from Cell Signaling Technology (Danvers, MA, USA).

Techniques: Phospho-proteomics, Protein-Protein interactions, Translocation Assay, Expressing, Migration

Journal: Oncology Letters

Article Title: Triptolide inhibits matrix metalloproteinase-9 expression and invasion of breast cancer cells through the inhibition of NF-κB and AP-1 signaling pathways

doi: 10.3892/ol.2021.12823

Figure Lengend Snippet: Effects of triptolide on TPA-induced NF-κB and AP-1 activation in MCF-7 cells. (A) MCF-7 cells were pretreated with inhibitors of NF-κB (Bay 11-7092) and AP-1 (SR 11302), and then TPA was added for 24 h. MMP-9 activity was detected by gelatin zymography and MMP-9 protein expression was analyzed by western blotting. (B) Cells were treated with triptolide and/or TPA. After a 3-h incubation, nuclear and cytoplasmic extracts were prepared. Translocation of p65, p50 and p-c-Fos to the nucleus and the levels of p-IκBα, p-IKKα/β, total c-Fos, IκBα, IKKα and IKKβ were determined by western blotting. PCNA was used as a loading control for the nucleus, and β-actin was used as an internal control for the cytoplasm. (C) NF-κB-luc and (D) AP-1-luc reporters were co-transfected with a Renilla luciferase thymidine kinase reporter into the cells. The cells were treated with TPA alone or with triptolide, and the NF-κB and AP-1 promoter activities were measured using a dual-luciferase reporter assay. The DNA binding of (E) NF-κB and (F) AP-1 was analyzed using electrophoretic mobility shift assays. Data are presented as the mean ± SEM of three independent experiments. *P<0.05 vs. TPA alone. AP-1, activator protein-1; MMP-9, matrix metalloproteinase-9; NF-κB, nuclear factor-κB; p-, phosphorylated; IKKα/β, IκB kinase α/β; IκBα, NF-κB inhibitor α; TPA, 12-O-tetradecanoylphorbol-13-acetate; zymo, zymography; PCNA, proliferating cell nuclear antigen.

Article Snippet: Rabbit antibodies against phosphorylated (p-)c-Fos (cat. no. 5348), p-IκB kinase α/β (p-IκKα/β; cat. no. 2697), stress activated protein kinase (SAPK)/JNK (cat. no. 9258), p-SAPK/JNK (cat. no. 4668), p38 MAPK (cat. no. 8690), p-p38 MAPK (cat. no. 4511), p44/42 MAPK (ERK1/2; cat. no. 4695), p-p44/42 MAPK (p-ERK1/2; cat. no. 4370) and c-Fos (cat. no. 2250) were purchased from Cell Signaling Technology, Inc. Rabbit antibodies against NF-κB p65 (cat. no. ab16502), NF-κB p105/p50 (cat. no. ab32360) and MMP-9 (cat. no. ab76003) were purchased from Abcam.

Techniques: Activation Assay, Activity Assay, Zymography, Expressing, Western Blot, Incubation, Translocation Assay, Control, Transfection, Luciferase, Reporter Assay, Binding Assay, Electrophoretic Mobility Shift Assay