HK-2 cells, a proximal tubular cell line derived from normal kidney, retain functional characteristics of proximal tubular epithelium. HK-2 cells were purchased from Procell Life Science & Technology Co., Ltd. (China), and cultured in Minimum Essential Medium (MEM, Procell) supplemented with 10% fetal bovine serum (TIANHANG, China) in a humidified 37°C incubator with 5% CO2. HK-2 cells were infected with the AV carrying shTOLLIP or TOLLIP and treated with 300 μM PQ for 24 h at 48 h postinfection at 37°C and 5% CO2. HK-2 cells were infected with the AV carrying shTOLLIP and treated with 300 μM PQ and 5 μM NF-κB inhibitor BAY11-7082 (Beyotime, China) for 24 h at 48 h postinfection at 37°C and 5% CO2.
Nf κb inhibitor bay 11 7082
Manufactured by Beyotime
Sourced in China
The NF-κB inhibitor Bay 11-7082 is a lab equipment product that functions as a potent and selective inhibitor of the NF-κB signaling pathway. It acts by blocking the phosphorylation of IκBα, a key regulatory protein in the NF-κB activation cascade.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Procell (1 mentions)
Hk 2 cells, by Procell (1 mentions)
Minimum essential medium (mem), by Procell (1 mentions)
Recombinant human tgf β1, by Thermo Fisher Scientific (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
Ckx41, by Olympus (1 mentions)
Anti phospho nf κb p65 antibody, by Cell Signaling Technology (1 mentions)
Dcfh da, by Merck Group (1 mentions)
Bca protein concentration assay kit, by Beyotime (1 mentions)
Irdye 800cw goat anti rabbit, by LI COR (1 mentions)
Anti cleaved notch1, by Cell Signaling Technology (1 mentions)
Anti p ikkα β, by Cell Signaling Technology (1 mentions)
Anti jag1, by Cell Signaling Technology (1 mentions)
Alexa fluor 488 goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
Anti p65, by Cell Signaling Technology (1 mentions)
Ly411575, by Merck Group (1 mentions)
Anti p iκbα, by Cell Signaling Technology (1 mentions)
Alexa fluor 555 goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions)
Anti acetyl histone h3, by Merck Group (1 mentions)
8 protocols using nf κb inhibitor bay 11 7082
1
Proximal Tubular Cell Knockdown and Oxidative Stress
2
Activated HSC Line LX-2 for In Vitro Studies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LX-2, an immortalized human HSC line with a stable phenotype and biochemical characteristics of activated HSCs42 (link), was verified by checking the ICLAC and NCBI databases and used for studies in vitro. LPS was purchased from Sigma-Aldrich (L2630, St. Louis, MO, U.S.). Recombinant human TGF-β1 was purchased from PeproTech (100–21C, Rocky Hill, U.S.). TLR4 inhibitor (TAK-242) was purchased from Calbiochem (614316, Darmstadt, Germany). NF-κB inhibitor (BAY11–7082) was purchased from Beyotime Biotechnology (S1523, Shanghai, China).
3
Cytokine Expression in BEAS-2B and HBEC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The expression of IL-8, IL-1β, and IL-6 were measured using an ELISA kit (Abcam) with detection limits of 1.8 pg/ml, 5.46 pg/ml, and 1.6 pg/ml, respectively. The levels of IL-8, IL-1β, and IL-6 were assayed by treatment of BEAS-2B cells and HBECs with recombinant HB-EGF. IL-8 depletion of cell supernatant was also confirmed by IL-8 ELISA assay. For experiments employing inhibitors, BEAS-2B cells and HBECs pretreated with 5 μM NF-κB inhibitor (Bay 11-7082, Beyotime Biotechnology) for 1 h and then treated with HB-EGF to assess the levels of IL-8.
4
Cell Invasion Assay with 25-HC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell invasion assay was carried out using 24-well cell culture chambers with an 8-µm pore filter (Corning Costar Corp., Cambridge, MA, USA). Before the cells were seeded, the upper surfaces of the membranes were coated with 50 µl Matrigel (BD Biosciences) for 6 h at 37°C. The cells were then seeded at a density of 1×105 cells/well in 100 µl serum-free F-12K medium added onto the top chamber of each Transwell with various concentrations of 25-HC. The cells were allowed to invade for 36 h at 37°C. After removing the cells that had remained in the upper side of the membrane with a cotton swab, cells in the lower side of the membrane were fixed with ice-cold methanol, stained with crystal violet (Hushi, Shanghai, China) in 20% ethanol overnight and then counted in 5 pre-determined fields under a light microscope (CKX41; Olympus).
For the inhibition assay, cells in the upper chamber were pre-treated with NF-κB inhibitor Bay 11-7082 at 2 µg/ml (Beyotime) for 1 h and the medium was replaced with FBS-free medium supplemented with 25-HC or not.
For the inhibition assay, cells in the upper chamber were pre-treated with NF-κB inhibitor Bay 11-7082 at 2 µg/ml (Beyotime) for 1 h and the medium was replaced with FBS-free medium supplemented with 25-HC or not.
5
Curcumol Modulates NF-κB Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Curcumol, curcumin (Chengdu Manste Biotechnology Co., Ltd.), DMEM high sugar culture liquid [Thermo Fisher biochemical products (Beijing) Co., Ltd.], fetal bovine serum (FBS) (Biological Industries), CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS), Red Tashan cigarette (Hongta Tobacco Co., Ltd.), SYBR Premix EX Tap II fluorescent quantitative reagent box (Takara Biological Engineering Co., Ltd.), Anti-Phospho-NF-κB p65 antibody (Cell Signaling Technology Company), Anti-Smad2+Smad3 (phosghoT8) antibody (Abcam Company), NF-κB inhibitor Bay 11-7082 (Beyotime Biotech Company), DCFH-DA (Sigma Company), and BCA protein concentration assay kit (Beyotime Biotech Company) were used in the study.
6
Signaling Pathway Profiling Using Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following primary antibodies were used: anti-RTA mouse monoclonal antibody was prepared in our laboratory; anti-Notch1 (Cell Signaling technology, #4380P), anti-cleaved Notch1 (Cell Signaling technology, #4147P), anti-JAG1 (Cell Signaling technology, 2620P), anti-HIF1-α (BD Biosciences, 610958), anti-acetyl-Histone H3 (Merck Millipore, 06–599), anti-H3K4me3 (Abcam, ab8580), anti-p-IKKα/β, anti-IKKα, anti-IKKβ, anti-IκBα, anti-p-IκBα, anti-p65 (Cell Signaling technology, 9958), anti-Flag (Sigma, F7425 and F1804), and anti-HA (Sigma, H9658 and H6908). The secondary antibodies were as follows: goat anti-mouse IRDye@800cw (LI-COR, 926–32210), goat anti-rabbit IRDye@800cw (LI-COR, 926–32211), goat anti-rabbit IgG(H+L) Alexa Fluor@488 (Invitrogen, R37116), and goat anti-mouse IgG(H+L) Alexa Fluor@555 (Invitrogen, A-21422). Other reagents used and their sources were as follows: anti-Flag M2 affinity gel (Sigma, A2220), DAPT (Sigma, D5942), LY-411575 (Sigma, SML0506), NF-κB inhibitor BAY 11–7082 (Beyotime, S1523), Wnt inhibitor Salinomycin (Sigma, S4526), and TNFα (Peprotech, 315-01A).
7
Icariin Enhances Osteoblast Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The α-modified Eagle’s medium–high glucose (α-MEM) was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Cell Proliferation ELISA (BrdU) kit was purchased from Roche (Mannheim, Germany). ALP vitality assay kit was purchased from Nanjing Jiancheng Biotechnology Co., Ltd. (Jiangsu, China). BCA Protein Assay Kit was obtained from Pierce (Rockford, IL, USA). Mammalian Cell Lysis Kit and UNIQ-10 column Trizol total RNA extraction kit were bought from Sangon Biological Engineering Technology and Services Co., Ltd. (Shanghai, China). Improm-II Reverse Transcription System was purchased from Promega Corporation (Madison, WI, USA). FastStart Universal SYBR Green Master (ROX) kit was purchased from Roche (Mannheim, Germany). P-IκB-α, IκB-α, p65, P-p65 monoclonal antibodies and peroxidase-conjugated secondary antibody were purchased from Cell Singaling Technology (USA), and β-actin monoclonal antibody was purchased from Sigma-Aldrich Co. (USA). NF-κB inhibitor BAY 11-7082 was purchased from Beyotime Institute of Biotechnology (China). Col I, OCN, BMP-2 monoclonal antibodies and peroxidase-conjugated secondary antibody were purchased from Santa Cruz Biotechnology (CA, USA). Icariin was purchased from the Chinese National Institute for Control of Pharmaceutical and Biological Products (Beijing, China).
8
Mesangial Cell Treatments and Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The MCs used in this study were from a mouse kidney mesangial cell line (SV40 MES-13, RRID: CVCL_5368), purchased from Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China). The MCs were cultured in the Dulbecco’s modified Eagle medium (DMEM, Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco). Different treatments of the cells were separately represented as follows: (1) The MCs were serum-starved overnight and exposed to LN serum (5% in DMEM) or control serum (5% in DMEM) for 24 h. The serum was detected for the endotoxin in advance and only serum with undetectable level of endotoxin could be used in this study. (2) The MCs were transfected with shRNA targeting DEC2 (Sh-DEC2) or negative control (Sh-NC). (3) The MCs were transfected with lentiviral vector overexpressing DEC2 (Lv-DEC2) or negative control (Lv-NC). (4) The MCs were treated with multiple inhibitors of signaling pathways, including TLR4-specific antagonist chloroquine phosphate (CP, MedChemExpress, Monmouth Junction, NJ, USA), PI3K inhibitor LY294002 (Beyotime Biotechnology), Akt inhibitor AZD5363 (Beyotime Biotechnology), NF-κB inhibitor BAY 11-7082 (Beyotime Biotechnology), p38 MAPK inhibitor SB203580 (Beyotime Biotechnology) and Erk inhibitor FR 180204 according to the manufacturer’s instructions.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!