phosphorylated p iκbα  (Santa Cruz Biotechnology)

Journal: Translational Cancer Research

Article Title: FXR activation suppresses NF-κB signaling, proliferation and migration in cervical cancer cells

doi: 10.21037/tcr-2025-522

Figure Lengend Snippet: FXR deficiency promotes cervical inflammation response. (A) Relative mRNA levels of pro-inflammatory genes in the cervix from the WT and FXR-KO mice, n=6. (B) Western blot showing p -IκBα, T-IκBα, p -STAT3, and T-STAT3 protein levels in the cervix from the WT and FXR-KO mice. β-actin served as a loading control. CXCL-2 , chemokine (C-X-C motif) ligand 2; FXR-KO, farnesoid X receptor-knockout; ICAM-1 , intercellular adhesion molecule 1; IL-1β , interleukin 1 beta; IL-6 , interleukin 6; IL-10 , interleukin 10; MCP-1 , chemokine (C-C motif) ligand 2; MMP2 , matrix metallopeptidase 2; MMP7 , matrix metallopeptidase 7; MMP10 , matrix metallopeptidase 10; mRNA, messenger RNA; p -IκBα, phosphorylated NF-κB inhibitor alpha; p -STAT3, phosphorylated STAT3; T-IκBα, NF-κB inhibitor alpha; T-STAT3, total STAT3; TNFα , tumor necrosis factor alpha; WT, wild-type.

Article Snippet: Anti-phosphorylated IκBα ( p -IκBα) and anti-IκBα (T-IκBα) antibodies were obtained from Cell Signaling Technologies (Danvers, MA, USA).

Techniques: Western Blot, Control, Knock-Out

Journal: Translational Cancer Research

Article Title: FXR activation suppresses NF-κB signaling, proliferation and migration in cervical cancer cells

doi: 10.21037/tcr-2025-522

Figure Lengend Snippet: Activation of FXR antagonizes the NF-κB signaling pathway in CC cells. (A) Hela and Siha cells were grown in 6-well plates for 16 hours and treated with 3 μM of GW4064 or DMSO for 24 hours, and then stimulated with TNFα (10 ng/mL) for another 1 hour. The cells were harvested, and the transcript levels of the NF-κB target genes were examined by qRT-PCR. (B) Hela cells were grown in 6-well plates for 16 hours, and transfected with p65 expression vector or control plasmid. After 24 hours of incubation, the cells were treated with 3 μM of GW4064 or DMSO for another 24 hours. The cells were harvested, and the transcript levels of the following cytokines were examined by qRT-PCR. (C) Relative luciferase activities of Hela cells that were co-transfected with the NF-κB reporter plasmid (pNF-κB-Luc), control phRL-TK plasmid, p65 expression plasmid, and FXR expression plasmids. The cells were treated with GW4064 (3 μM) or DMSO for 24 hours. (D,E) Representative immunoblots showing p -IκBα, T-IκBα, P-STAT3, and T-STAT3 protein levels in the Hela (D) and Siha (E) cells treated with DMSO, TNFα (10 ng/mL), or IL-6 (10 ng/mL), GW4064 (3 μM), TNFα + GW4064. β-actin served as a loading control. *, P<0.05. CC, cervical cancer; CON, control; DMSO, dimethyl sulfoxide; FXR , farnesoid X receptor; ICAM-1 , intercellular adhesion molecule 1; IL-1α , interleukin 1 alpha; IL-1β , interleukin 1 beta; IL-6 , interleukin 6; INOS , inducible nitric oxide synthase; IP-10 , interferon-inducible protein-10; MCP-1 , chemokine (C-C motif) ligand 2; MMP2 , matrix metallopeptidase 2; mRNA, messenger RNA; NF-κB, nuclear factor kappa B; p -IκBα, phosphorylated NF-κB inhibitor alpha; p -STAT3, phosphorylated STAT3; qRT-PCR, quantitative real-time polymerase chain reaction; RLU, relative light unit; T-IκBα, NF-κB inhibitor alpha; T-STAT3, total STAT3; TGFβ2 , transforming growth factor beta 2; TNFα , tumor necrosis factor alpha.

Article Snippet: Anti-phosphorylated IκBα ( p -IκBα) and anti-IκBα (T-IκBα) antibodies were obtained from Cell Signaling Technologies (Danvers, MA, USA).

Techniques: Activation Assay, Quantitative RT-PCR, Transfection, Expressing, Plasmid Preparation, Control, Incubation, Luciferase, Western Blot, Real-time Polymerase Chain Reaction

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fibroblast Growth Factor 20 Attenuates Colitis by Restoring Impaired Intestinal Epithelial Barrier Integrity and Modulating Macrophage Polarization via S100A9 in an NF-κB-Dependent Manner

doi: 10.1016/j.jcmgh.2025.101486

Figure Lengend Snippet: FGF20 inhibits NF-κB pathway through S100A9 in DDS-induced mice. ( A ) KEGG Enrichment Analysis of proteomics data from the colonic tissue of FGF20 KO and WT mice with or without colitis. ( B ) Immunoblot analysis of colonic p-P65, P65, p-IκBα, and IκBα from FGF20 KO and WT mice with or without colitis (n = 6). ( C ) Immunoblot analysis of colonic p-P65, P65, p-IκBα, and IκBα contents in AAV-FGF20- or AAV-GFP-treated mice with or without colitis. ( D ) Immunoblot analysis of colonic p-P65, P65, p-IκBα, and IκBα contents in FGF20 KO mice and WT mice with colitis, treated with paquinimod or vehicle (n = 6). ( E ) Immunoblot analysis of colonic p-P65, P65, p-IκBα, and IκBα contents in AAV-FGF20-treated mice and AAV-GFP-treated mice with colitis, treated with AAV-S100A9 or not (n = 6). Data are represented as mean ± SEM; ∗ P < .05, ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: FGF20 (1/500, SC-373927, Santa CruzA), Bcl2 (1/1000, AB182858, Abcam), Bax (1/1000, AB32503, Abcam), S100A9 (1/500, SC-53187, Santa Cruz), S100A8 (1/500, SC-48352, Santa Cruz), phosphorylated (p) -IκBα (1/500, SC-8404, Santa Cruz), p-P65 (1/500, SC-136548, Santa Cruz), IκBα (1/1000, 4814S, CST, USA), P65 (1/1000, 4814S, CST), TNF-α (1/1000, AF7014, Affinity, China), IL6 (1/1000, 12912S, CST), FGFR1 (1/1000, bs-0230R, Bioss, China), FGFR2 (1/1000, bs-0675R, Bioss), FGFR3 (1/2000, MA5-31142, ThermoFisher), FGFR4 (1/1000, bsm-61509R, Bioss), GAPDH (1/10000, HRP-60004, Proteintech, China), and Beta-Tubulin (1/5000, 10094-1-AP, Proteintech).

Techniques: Western Blot

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Fibroblast Growth Factor 20 Attenuates Colitis by Restoring Impaired Intestinal Epithelial Barrier Integrity and Modulating Macrophage Polarization via S100A9 in an NF-κB-Dependent Manner

doi: 10.1016/j.jcmgh.2025.101486

Figure Lengend Snippet: FGF20 inhibits M1-like macrophage polarization in NF-κB-dependent manner in vitro. Primary peritoneal macrophages were isolated from WT mice and FGF20 KO mice and used for subsequent in vitro experiments. ( A ) Immunoblot analysis of expression of iNOS, TNF-α, IL6, S100A9, p-P65, P65, p-IκBα, and IκBα in LPS-stimulated primary peritoneal macrophages, treated with or without BAY11-7082 (n = 3). ( B ) Representative immunofluorescence images and its fluorescence densities of LPS-stimulated primary peritoneal macrophages, treated with or without BAY11-7082 (n = 6). Data are represented as mean ± SEM; ∗∗ P < .01, ∗∗∗ P < .001.

Article Snippet: FGF20 (1/500, SC-373927, Santa CruzA), Bcl2 (1/1000, AB182858, Abcam), Bax (1/1000, AB32503, Abcam), S100A9 (1/500, SC-53187, Santa Cruz), S100A8 (1/500, SC-48352, Santa Cruz), phosphorylated (p) -IκBα (1/500, SC-8404, Santa Cruz), p-P65 (1/500, SC-136548, Santa Cruz), IκBα (1/1000, 4814S, CST, USA), P65 (1/1000, 4814S, CST), TNF-α (1/1000, AF7014, Affinity, China), IL6 (1/1000, 12912S, CST), FGFR1 (1/1000, bs-0230R, Bioss, China), FGFR2 (1/1000, bs-0675R, Bioss), FGFR3 (1/2000, MA5-31142, ThermoFisher), FGFR4 (1/1000, bsm-61509R, Bioss), GAPDH (1/10000, HRP-60004, Proteintech, China), and Beta-Tubulin (1/5000, 10094-1-AP, Proteintech).

Techniques: In Vitro, Isolation, Western Blot, Expressing, Immunofluorescence, Fluorescence

Journal: Immunity, inflammation and disease

Article Title: Pirfenidone alleviates smoke inhalation lung injury of rats via the NF-κB signaling pathway.

doi: 10.1002/iid3.70014

Figure Lengend Snippet: FIGURE 3 PFD suppressed the activation of NF‐κB pathway in SILI rats. (A) Western blot was adopted to elucidate the levels of p‐IκBα and p‐p65 in lung tissues of SILI rats. (B) Western blot was adopted to elucidate the levels of p‐IκBα and p‐p65 in lung tissues of SILI rats after addition of NF‐κB agonist PMA. N = 5. Results are presented as means ± SD, analyzed using ANOVA, followed by Turkey post hoc test. ***p < .001 versus Control group; ##p < .01, ###p < .001 versus SILI group; &p < .05 versus SILI + PFD group. ANOVA, analysis of variance; NF‐κB, nuclear factor kappa B; SD, standard deviation; SILI, smoke inhalation lung injury.

Article Snippet: After blocked with 5% nonfat milk, the membranes were incubated with primary antibodies against phosphorylation‐ IκBα (p‐IκBα) (#2859, 1:1000, Cell Signaling Technology), IκBα (#4812, 1:1000, Cell Signaling Technology), p‐p65 (ab76302, 1:1000, Abcam), p65 (ab16502, 1:1000, Abcam), and GAPDH (ab181602, 1:2000, Abcam) overnight at 4°C, followed by horseradish peroxidase‐conjugated goat anti‐ rabbit IgG secondary antibody (ab205718, 1:2000, Abcam) at 37°C for 1 h. The protein blots were visualized with the enhanced chemiluminescence kit and the optical density was analyzed with ImageJ software (National Institute of Health, Bethesda).

Techniques: Activation Assay, Western Blot, Control, Standard Deviation