Journal: International journal of radiation oncology, biology, physics

Article Title: Therapeutic Intervention Using a Smad7-Based Tat Protein to Treat Radiation-Induced Oral Mucositis.

doi: 10.1016/j.ijrobp.2021.09.039

Figure Lengend Snippet: Fig. 5. Pharmacodynamics markers of Tat-PYC-Smad7 treated dog oral mucositis. (A) Biopsies of posttreatment oral mucosa were evaluated by immunohistochemistry to detect NFkB-p50, pSmad3, pH2AX, and myeloperoxidase. Representative images of vehicle treated and Tat-PYC-Smad7 treated mucosa are presented with the lower panel in each set, presenting a high-power view of the blue box identified in the panel above. Scale bar: 100 mm. (B) Quantification of positive, brown-staining in epithelium sections excluding ulcers (in vehicle group) pictured in panel A. Myeloperoxidase staining together with mor- phology evaluation were used to identify neutrophils (indicated by yellow arrows) in ulcer surface and stroma. Faint brown staining in epithelium is nonspecific. The staining quantification for each dog and the mean of each treatment § standard error of the mean is presented.

Article Snippet: We performed immunohistochemistry (IHC) staining as previously described.15 Primary antibodies used were rabbit anti-Ki67 (1:500) (Cell Signaling Technology, 12202), rabbit anti-NFkB-p105/p50 [p Ser337] (1:300) (Novus Biologicals, NB100-82074), rabbit anti-pSmad3 (1:400, Abcam, ab52903), rabbit anti-pH2AX (1:100) (Cell Signaling Technology, 9718), rabbit anti-myeloperoxidase (Ready-to-Use, Dako, GA51161-2), rabbit anti-hemagglutinin (HA) (1:700) (Cell Signaling Technology, 3724), rabbit anti-IL-1b (1:500) (Novus Biologicals, NB600-633), rabbit anti-TNF-a (1:200) (Novus Biologicals, NBP1-19532), mouse anti-CCL2 (1:50) (Novus Biologicals, MAB28171), and goat anti-IL-6 (1:40) (R&D, AF1609) antibodies.

Techniques: Drug discovery, Immunohistochemistry, Staining

Journal: eLife

Article Title: Single-PanIN-seq unveils that ARID1A deficiency promotes pancreatic tumorigenesis by attenuating KRAS -induced senescence

doi: 10.7554/eLife.64204

Figure Lengend Snippet:

Article Snippet: Antibody , (Rabbit polyclonal) anti-Ras , Abcam , Abcam Cat# ab180772, RRID: AB_2884935 , WB(1:500).

Techniques: Recombinant, Expressing, shRNA, Sequencing, Detection Assay, Staining

Journal: Cell Death & Disease

Article Title: CCL11 promotes hepatocellular carcinoma recurrence after surgery by potentiating immunosuppressive CCR5 + CD206 + M2-like macrophages and promoting tumor invasiveness

doi: 10.1038/s41419-026-08508-4

Figure Lengend Snippet: A Flow cytometry quantification of the number of monocyte-differentiated CCR5 + CD206 + M2-like macrophages after administration of recombinant human CCL11 (rhCCL11) protein. Control group: PBS treatment. B The expression Levels of M2-macrophage-related immunosuppressive markers and cytokines in CCL11-treated and PBS-treated CCR5 + M2-like macrophages. C Cytokine array analysis of secreted cytokines in CCL11-treated and PBS-treated CCR5 + M2-like macrophages. Red dot boxes indicate two highly upregulated cytokines: 1, MCP1; 2, IL-8. D mRNA Levels of IL-8 and MCP-1 in CCL11-treated and PBS-treated CCR5 + M2-like macrophages. E CCL11-treated CCR5 + M2-like macrophages enhanced the induction of CD4+ Foxp3+Tregs from naive CD4 + T compared to the PBS-treated control group. F Top 10 enriched KEGG pathways from differentially expressed genes in the CCL11-treated CCR5 + M2-like macrophages. G Gene set enrichment analysis of Toll-like receptor signaling pathway. H Levels of p-IKKα/β, p-IkBα, NFkB1-P50, and PD-L1 expression via western blot analysis. I Schematic diagram of the molecular mechanism of CCL11 in regulating CCR5 + M2-like macrophages. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns not significant.

Article Snippet: Antibodies used for Western blot analysis anti-human/mouse p-IKKα/β (#2078T), anti-human/mouse p-IkBα (#2859T), anti-human/mouse NFKB1-p50 (#13586S), anti-human/mouse p-AKT (#9271S), and anti-human/mouse AKT (#9272S) from Cell Signaling Technology; anti-human/mouse PDL1 (#14-5983-82), anti-human/mouse p-PI3K (#PA5-104853), and anti-human/mouse Mafk (#PA5-115534) from Thermo Fisher Scientific; anti-human/mouse PI3K (#sc-423) and anti-human GAPDH (#sc47724) from Santa Cruz Biotechnology; and anti-human/mouse MMP13 (#18165-1-AP) and anti-mouse GAPDH (#60004-1-Ig) from Proteintech Group.

Techniques: Flow Cytometry, Recombinant, Control, Expressing, Western Blot

Journal: Molecular Medicine Reports

Article Title: Downregulation of transgelin 2 promotes breast cancer metastasis by activating the reactive oxygen species/nuclear factor-κB signaling pathway

doi: 10.3892/mmr.2019.10643

Figure Lengend Snippet: Loss of TAGLN2 expression induces NF-κB activation. (A) Luciferase activity of an NF-κB-luciferase reporter plasmid in TAGLN2-overexpressing 293T cells was measured 48 h after stimulation with siRNA-TAGLN2 or scramble siRNA; luciferase activity was normalized to that of Renilla . Data were representative of three independent experiments. (B) Immunoblotting of IκB, p50, p65/RelA and TAGLN2 expression in MDA-MB-231 and MCF-7 cells transfected with pLKO.1-shRNA-TAGLN2 and pLKO.1-shRNA-scramble. The results of western blotting were representative of three independent experiments. (C) Cell migration was promoted in MDA-MB-231 cells transfected with pLKO.1-shRNA-TAGLN2 compared with cells treated with scramble shRNA. Treatment with 18 µM SN50 for 24 h reversed the effects of TAGLN2 downregulation on cell invasion. Data are representative of three independent experiments. *P<0.05 vs. scramble or as indicated. DMSO, dimethyl sulfoxide; NF-κB, nuclear factor-κB; shRNA, short hairpin RNA; siRNA, small interfering RNA; TAGLN2, transgelin 2.

Article Snippet: The membranes were probed with primary antibodies against PRDX1 (1:1,000; cat. no. EPR5433; Epitomics, Abcam, Cambridge, UK), FLAG (1:500; cat. no. F7425; Sigma-Aldrich, Merck KGaA), TAGLN2 (1:1,000; cat. no. 60044-1-lg; ProteinTech Group, Inc.), IκB (1:3,000; cat. no. 66418-1-lg; ProteinTech Group, Inc.), p50 (1:200; cat. no. 15506-1-AP; ProteinTech Group, Inc.), p65 (1:1,000; cat. no 10745-1-AP; ProteinTech Group, Inc.), MMP1 (1:1,000; cat. no. 54376; Cell Signaling Technology), MMP2 (1:1,000; cat. no. 4022; Cell Signaling Technology), CXCR4 (1:1,000; cat. no. ab181020; Abcam, Cambridge, UK), Lamin B1 (1:3,000; cat. no. 12987-1-AP; ProteinTech Group, Inc.) and GAPDH (1:3,000; cat. no. 60004-1-lg; ProteinTech Group, Inc.) on the rocking table at 4°C overnight.

Techniques: Expressing, Activation Assay, Luciferase, Activity Assay, Plasmid Preparation, Western Blot, Transfection, shRNA, Migration, Small Interfering RNA

Journal: The Indonesian Biomedical Journal

Article Title: Zinc Administration Affects Bronchial Mucosal NF-κB p105/p50, p-NF-κB p65, IL-8, and IL-1β of Zinc-deficient Rats

doi: 10.18585/inabj.v12i3.1041

Figure Lengend Snippet: Figure 1. Immuno-expression of NF-κB p105/p50 in bronchial mucosae. A: Z1 group, B: Z2 group, C: Z3 group, D: Z4 group. Black bar: 10 µm.

Article Snippet: For NF-κB p105/p50 detection, Results a mouse monoclonal anti-NF-κB p105/p50 antibody (Cat# NB100-56583, Novus Biologicals, Centennial, CO, USA) was applied.

Techniques: Expressing

Journal: The Indonesian Biomedical Journal

Article Title: Zinc Administration Affects Bronchial Mucosal NF-κB p105/p50, p-NF-κB p65, IL-8, and IL-1β of Zinc-deficient Rats

doi: 10.18585/inabj.v12i3.1041

Figure Lengend Snippet: Figure 2. Immuno-expression of p-NF-κB p65 in bronchial mucosae. A: Z1 group, B: Z2 group, C: Z3 group, D: Z4 group. Black bar: 10 µm.

Article Snippet: For NF-κB p105/p50 detection, Results a mouse monoclonal anti-NF-κB p105/p50 antibody (Cat# NB100-56583, Novus Biologicals, Centennial, CO, USA) was applied.

Techniques: Expressing

Journal: eLife

Article Title: PTH regulates osteogenesis and suppresses adipogenesis through Zfp467 in a feed-forward, PTH1R-cyclic AMP-dependent manner

doi: 10.7554/eLife.83345

Figure Lengend Snippet: ( A ) qPCR results of overexpression of Ebf1 , Myod , Myog , Gata1, and NFκB1 in MC3T3-E1 cell line. GATA1 and NFκB1 overexpression could significantly upregulate the expression level of Pth1r . Data shown as mean ± SD by unpaired Student’s t test, n=3 independent experiments for each group. ( B ) Representative confocal images of GATA1 and NFκB1 immunofluorescence in Zfp467 +/+ and Zfp467 -/- bone marrow stromal cells (BMSCs) and related quantification. ( C ) Nuclear protein level of GATA1 and NFκB1 in Zfp467 +/+ and Zfp467 -/- BMSCs. ( D ) Quantification analysis for nuclear protein level of GATA1 and NFκB1 in Zfp467 +/+ and Zfp467 -/- BMSCs. Data shown as mean ± SD, n=3 independent experiments for each group. Data shown as mean ± SD by unpaired Student’s t test, n=3 independent experiments for each group. Figure 4—source data 1. Western blot for . Figure 4—source data 2. Western blot for ACTIN. Figure 4—source data 3. Western blot for H3. Figure 4—source data 4. Western blot for p50 in cytoplasm. Figure 4—source data 5. Western blot for GATA1 in cytoplasm. Figure 4—source data 6. Western blot for GATA1 in nuclear. Figure 4—source data 7. Western blot for p50 in nuclear.

Article Snippet: NFκB1 (IB) , Cell Signaling Technology , 13681S.

Techniques: Over Expression, Expressing, Immunofluorescence, Western Blot

Journal: eLife

Article Title: PTH regulates osteogenesis and suppresses adipogenesis through Zfp467 in a feed-forward, PTH1R-cyclic AMP-dependent manner

doi: 10.7554/eLife.83345

Figure Lengend Snippet: ( A ) Reporter assays using the indicated P1 or P2 reporter construct and an expression vector bearing Gata1 , Nfkb1, or a control empty vector. Data shown as mean ± SD by one-way ANOVA, n=3 independent experiments for each group. Data shown as mean ± SD by one-way ANOVA, n=3 independent experiments for each group. ( B ) Immunoblot assay using a control rabbit IgG antibody (IgG) or the anti-NFκB1 antibody during chromatin immunoprecipitation assay. ( C ) DNA enrichment of Pth1r P2 promoter, ratio between NFκB1 and IgG IP products, first part and last two parts of P2 were significantly enriched by NFκB1 antibody. Data shown as mean ± SD by unpaired Student’s t test, n=3 independent experiments for each group. ( D, E ) qPCR results of the expression levels of Nfkb1 and Pth1r in Nfkb1 siRNA-treated Zfp46 +/+ and Zfp467 -/- calvarial osteoblasts (COBs) and bone marrow stromal cells (BMSCs). Data shown as mean ± SD by two-way ANOVA, n=3 independent experiments for each group. ( F ) Western blot analysis of Nfkb1 and Pth1r in Nfkb1 siRNA-treated Zfp467 +/+ and Zfp467 -/- COBs and BMSCs. ( G ) Quantification for PTH1R protein level. Data shown as mean ± SD by two-way ANOVA, n=3 independent experiments for each group. NC, negative control. Figure 5—source data 1. Western blot for . Figure 5—source data 2. Western blot for p50 with IP samples. Figure 5—source data 3. Western blot for PTH1R. Figure 5—source data 4. Western blot for p50. Figure 5—source data 5. Western blot for ACTIN.

Article Snippet: NFκB1 (IB) , Cell Signaling Technology , 13681S.

Techniques: Construct, Expressing, Plasmid Preparation, Control, Western Blot, Chromatin Immunoprecipitation, Negative Control

Journal: eLife

Article Title: PTH regulates osteogenesis and suppresses adipogenesis through Zfp467 in a feed-forward, PTH1R-cyclic AMP-dependent manner

doi: 10.7554/eLife.83345

Figure Lengend Snippet: ( A ) DNA enrichment of Pth1r P2 promoter, ratio between IP and input, first two parts of P2 were enriched using NFκB1 antibody. Data shown as mean ± SD by unpaired Student’s t test, n=3 replicates for each group. ( B ) RT-PCR product of chromatin immunoprecipitation assay using the nuclear extracts from MC3T3-E1 cells. Figure 5—figure supplement 1—source data 1. Nucleic acid blot for . Figure 5—figure supplement 1—source data 2. Nucleic acid blot for , PCR blot for Pth1r amplification product.

Article Snippet: NFκB1 (IB) , Cell Signaling Technology , 13681S.

Techniques: Reverse Transcription Polymerase Chain Reaction, Chromatin Immunoprecipitation, Amplification

Journal: eLife

Article Title: PTH regulates osteogenesis and suppresses adipogenesis through Zfp467 in a feed-forward, PTH1R-cyclic AMP-dependent manner

doi: 10.7554/eLife.83345

Figure Lengend Snippet: ( A ) DNA pulldown assay with biotin-labeled Pth1r P2. MC3T3-E1 nuclear extracts or NFκB1 recombinant protein was probed with biotin-Pth1rP2 and then subjected to immunoblotting using NFκB1 antibody. ( B ) qPCR results of the expression levels of Nfyc , Npas1, Rel , Akap8 , Rela , Ankrd42 , Relb, and Hdac1 in related siRNA-treated MC3T3-E1 cells. Data shown as mean ± SD by one-way ANOVA, n=3 independent experiments for each group. NC, negative control. ( C ) qPCR results of the expression levels of Pth1r in Nfkb1 overexpression plasmid and Nfyc , Npas1, Rel , Akap8 , Rela , Ankrd42 , Relb , or Hdac1 siRNA co-transfected MC3T3-E1 cells. NC, negative control. ( D ) IP results using NFκB1 antibody in MC3T3-E1 protein extracts, IgG was used as a negative control. ( E ) Protein level of p-p105 and p50 in Zfp467 knockdown MC3T3-E1 cells. Data shown as mean ± SD by unpaired Student’s t test, n=3 independent experiments for each group. ( F ) Protein and mRNA level of NFκB-inducing kinase (NIK) in control and Zfp467 siRNA-treated MC3T3-E1 cells. Data shown as mean ± SD by unpaired Student’s t test, n=3 independent experiments for each group. ( G ) Protein level of p-IKKa and p-p100 and mRNA level of Ikka in control and Zfp467 siRNA-treated MC3T3-E1 cells. Data shown as mean ± SD by unpaired Student’s t test, n=3 independent experiments for each group. Figure 6—source data 1. Western blot for . Figure 6—source data 2. Western blot for p50 in DNA pulldown experiment using nuclear extract. Figure 6—source data 3. Western blot for p50 in DNA pulldown experiment using p50 purified protein. Figure 6—source data 4. Western blot for RelB. Figure 6—source data 5. Western blot for NPAS1. Figure 6—source data 6. Western blot for p50. Figure 6—source data 7. Western blot for p-p105. Figure 6—source data 8. Western blot for p50/p105. Figure 6—source data 9. Western blot for ACTIN. Figure 6—source data 10. Western blot for NFκB-inducing kinase (NIK). Figure 6—source data 11. Western blot for ACTIN. Figure 6—source data 12. Western blot for p-IKKα/β. Figure 6—source data 13. Western blot for IKKα. Figure 6—source data 14. Western blot for p-p100. Figure 6—source data 15. Western blot for p100.

Article Snippet: NFκB1 (IB) , Cell Signaling Technology , 13681S.

Techniques: Labeling, Recombinant, Western Blot, Expressing, Negative Control, Over Expression, Plasmid Preparation, Transfection, Knockdown, Control, Purification

Journal: eLife

Article Title: PTH regulates osteogenesis and suppresses adipogenesis through Zfp467 in a feed-forward, PTH1R-cyclic AMP-dependent manner

doi: 10.7554/eLife.83345

Figure Lengend Snippet: Antibody list for co-IP, ChIP, and western blot.

Article Snippet: NFκB1 (IB) , Cell Signaling Technology , 13681S.

Techniques: Western Blot

Journal: Cell Reports Medicine

Article Title: Oncostatin M signaling drives cancer-associated skeletal muscle wasting

doi: 10.1016/j.xcrm.2024.101498

Figure Lengend Snippet:

Article Snippet: NF-κB1 p105/p50 (D7H5M) Rabbit Antibody , Cell Signaling , Cat# 12540; RRID: AB_2687614.

Techniques: Ubiquitin Proteomics, Virus, Recombinant, SYBR Green Assay, Transfection, Expressing, Purification, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software