The MEK1/2/ERK1/2-specific inhibitor U0126 (catalog no. s1901; Beyotime) was used to inhibit the activation of MEK1/2 (32 (link)). PK-15 cells were inoculated with CSFV as described above. At −2, 0, or 2 hpi, U0126 was added to the cells cultured in serum-free DMEM at a final concentration of 30 μM. The MEK1/ERK1/2-specific inhibitor PD98059 (catalog no. s1805; Beyotime) was used as a negative control to inhibit the activation of MEK1 (32 (link)). At 72 hpi, the supernatants were collected to detect the yields of CSFV progeny virus and viral genome copy numbers, and the cells were lysed with 100 μl of ice-cold radioimmunoprecipitation assay (RIPA) lysis buffer (catalog no. P0013C; Beyotime) containing complete protease inhibitor cocktail (catalog no. 11697498001; Roche) and phosphatase inhibitor cocktail (catalog no. 04906845001; Roche) for Western blotting.
U0126
Manufactured by Beyotime
Sourced in China, United States
U0126 is a selective inhibitor of the mitogen-activated protein kinase kinase (MEK1 and MEK2) enzymes. It is a commonly used tool in cell biology research to study the role of the ERK/MAPK signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Sp600125, by Beyotime (19 mentions)
Sb203580, by Beyotime (17 mentions)
Ly294002, by Beyotime (11 mentions)
Bay 11 7082, by Beyotime (7 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (5 mentions)
β actin, by Santa Cruz Biotechnology (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
P jnk, by Beyotime (3 mentions)
Pd98059, by Beyotime (3 mentions)
Sb203580, by Merck Group (2 mentions)
Ly294002, by Selleck Chemicals (2 mentions)
Dimethyl sulfoxide (dmso), by Beyotime (2 mentions)
Epidermal growth factor (egf), by R&D Systems (2 mentions)
F 12k medium, by Thermo Fisher Scientific (2 mentions)
Vimentin, by Cell Signaling Technology (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Ribavirin, by Merck Group (2 mentions)
Gapdh, by Santa Cruz Biotechnology (2 mentions)
Alizarin red s, by Merck Group (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
55 protocols using u0126
1
Inhibition of CSFV Replication by U0126
2
Signaling Pathways in H2O2-Induced Granulosa Cell Responses
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H89, LY294002, SB203580 and U0126 were purchased from Beyotime (Beijing, China). After exposure to 200 μM H2O2 (Sigma, St. Louis, MO, USA) for 1 h, MGCs were rinsed with PBS and grown in serum-free DMEM/F-12 containing 7.5 IU/ml FSH for 2, 6, 12, 24 or 36 h. In some experiments, H89 (10 μM), LY294002 (20 μM), SB203580 (20 μM) or U0126 (3 μM) was added 30 min before FSH treatment.
3
Reagents and Antibodies for Cell Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Reagents. Neutral red staining solution was obtained from Sangon Biotech (Shanghai, China). U0126 (MEK1/2 inhibitor), SP600125 (JNK inhibitor) and SB203580 (p38 inhibitor) were purchased from Beyotime (Shanghai, China). LY294002 (PI3K inhibitor) was purchased from Selleck Chemicals (Houston, TX, USA). Anti-FIP-glu antiserum was raised in rabbits (99) . Anti-6×His Tag mouse monoclonal antibody, HRP-conjugated Goat Anti-Mouse IgG and HRP-conjugated Goat Anti-Rabbit IgG were from Sangon Biotech (Shanghai, China).
4
LPS-Induced Inflammatory Response in RAW 264.7 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RAW 264.7 cells were cultured in 6-well culture dishes until they reached approximately 80% confluence and were then stimulated with 1 μg/mL Escherichia coli LPS (InvivoGen, San Diego, CA, USA) for a certain number of hours. Cells not stimulated by LPS were used as a control.
In the signaling pathway inhibition experiments, the cells were first treated with the signaling pathway inhibitor BAY 11-7082 (Beyotime, Shanghai, China; 10 μM), U0126 (Beyotime, Shanghai, China; 10 μM), SB203580 (Beyotime, Shanghai, China; 20 μM) or SP600125 (Beyotime, Shanghai, China; 20 μM) for 1 h, and were then stimulated with 1 μg/mL LPS for 6 h. Cells not stimulated with LPS or treated with signaling pathway inhibitors were used as a blank control.
In the signaling pathway inhibition experiments, the cells were first treated with the signaling pathway inhibitor BAY 11-7082 (Beyotime, Shanghai, China; 10 μM), U0126 (Beyotime, Shanghai, China; 10 μM), SB203580 (Beyotime, Shanghai, China; 20 μM) or SP600125 (Beyotime, Shanghai, China; 20 μM) for 1 h, and were then stimulated with 1 μg/mL LPS for 6 h. Cells not stimulated with LPS or treated with signaling pathway inhibitors were used as a blank control.
5
Bovine IL-1α Signaling Pathway Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Recombinant bovine IL-1α was purchased from Kingfisher Biotech. U0126, SP600125, SB203580 and BAY11-7082 were obtained from Beyotime Biotechnology. Erk antibody, p-Erk antibody, c-Jun antibody, p-c-Jun antibody, IκBα antibody and p-IκBα antibody were purchased from Cell Signaling Technology. p38 antibody, p-p38 antibody and β-actin antibody were purchased from LifeSpan BioSciences. Forskolin were purchased from Abcam.
6
ApoE Isoform Effects on Axonal Regeneration
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Three human recombinant ApoE proteins, isoforms 2, 3 and 4 (PeproTech, Rocky Hill, NJ, USA), were separately added into Neurobasal/B27 to culture the explants of ApoE-/- mice after planting. The ApoE final concentration was 10 µg/ml in medium, which is similar to the concentration of ApoE in human cerebrospinal fluid (CSF)22 (link)–24 (link, link) . In addition, after axonal transection, the explants of the ApoE-/- mice were continuously cultured in Neurobasal/B27 with human recombinant ApoE2/3/4 proteins for 24 h.
JNK/ERK/p38 inhibitors, which included SP600125, U0126, and SB203580 (Beyotime, Shanghai, China), were also used as interventional means. The final concentration of the three inhibitors was 10 μM, and they were dissolved in dimethyl sulfoxide (DMSO, Beyotime)25 (link),26 (link). In the control group, only DMSO was added without the inhibitors. After explant planting, the explants were separately treated by medium with the JNK, ERK and p38 inhibitors for 24 h. Subsequently, the media were replaced with fresh media without the inhibitors27 (link).
JNK/ERK/p38 inhibitors, which included SP600125, U0126, and SB203580 (Beyotime, Shanghai, China), were also used as interventional means. The final concentration of the three inhibitors was 10 μM, and they were dissolved in dimethyl sulfoxide (DMSO, Beyotime)25 (link),26 (link). In the control group, only DMSO was added without the inhibitors. After explant planting, the explants were separately treated by medium with the JNK, ERK and p38 inhibitors for 24 h. Subsequently, the media were replaced with fresh media without the inhibitors27 (link).
7
Preparation and Storage of Common Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
U0126 was purchased from Beyotime (Shanghai, China). All other compounds used in this study were purchased from Selleckchem (Houston, USA). All the compounds were dissolved at 10 mM in DMSO and stored at -20°C. IGF, FGFα and HGF were purchased from Prepro Tech (New Jersey Rocky Hill, USA); NRG1 and EGF were purchased from R&D System (Minneapolis, MN, USA) and Sigma-Aldrich (St. Louis, MO, USA), respectively. All growth factors were dissolved in 0.1% BAS as stock solution and the aliquots were stored at -20°C.
8
SV-HUC-1 Cell Line Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An SV-40 immortalized human urothelial cell line (SV-HUC-1) was purchased from Chinese Academy of Typical Culture Collection Cell Bank. F12K medium was purchased from Gibco (New York, NY, USA). Fetal bovine serum (FBS) was obtained from PAA Laboratories (Pasching, Austria). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was purchased from Sigma-Aldrich. SB203580, SP600125, and U0126 were purchased from Beyotime (Shanghai, China). The primary antibodies for phosphorylated JNK, phosphorylated p38, phosphorylated ERK1/2, phosphorylated c-Jun, phosphorylated c-Fos, E-cadherin, ZO-1, N-cadherin and Vimentin were obtained from Cell Signaling Technology (Beverly, MA). GAPDH antibody was from Biogot Technology (Nanjing, China). Primers for E-cadherin, ZO-1, Vimentin, N-cadherin and GAPDH were synthesized by Invitrogen (Carlsbad, CA). Sources of other materials are noted accordingly in the text.
9
Cell Line Culture and Reagent Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell lines including SKOV3, OVCAR3, HeLa, SiHa, GLC-82, A549 and HepG2 were purchased from American Type Culture Collection (ATCC) and were cultured in Dulbecco's Modified Eagle Medium (DMEM), supplemented with 10% fetal bovine serum (FBS), streptomycin (100 mg/mL) and penicillin (100 U/mL) at 37°C in a 5% CO2 humid incubator. DCA and Cycloheximide (CHX) were purchased from Sigma-Aldrich (Louis, MO, USA). Met, U0126, MG132 and Hoechst 33258 were purchased from Beyotime Company (Shanghai, China). MK2206 was purchased from Selleck Company (Shanghai, China). Caspase-3 Activity Assay Kit and Reactive Oxygen Species Assay Kit were purchased from Beyotime Company (Shanghai, China). Annexin V-FITC and PI were purchased from BD Bioscience (BD, NJ, USA). siRNAs to Mcl-1, ATG7, PDH and control siRNA were from GenePharma (Shanghai, China). pCMV and pcDNA3.1, pCMV-Mcl-1, pcDNA3.1-PDK1 and pcDNA3.1-PDK2 expression plasmids were bought from Obio Technology (Shanghai, China).
10
Quantification of Secreted Cytokines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Secreted human IL‐1β and IL‐6 in culture supernatants were quantified using ELISA kits (Thermo Scientific) according to the manufacturer's instructions. LY294002 (phosphatidylinositol 3‐kinase, PI3K inhibitor) and U0126 (Erk inhibitor) were obtained from Beyotime Biotechnology (Shanghai, China). Experiments were performed in triplicate.
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!