PJ34 was purchased from Enzo Life Sciences (Farmingdale, NY, USA); ABT888 from APExBIO (Houston, TX, USA); XAV939 from Cayman chemical company (Ann Arbor, MI, USA); MG-132, ICG-001, and PNU-74654 from Selleckchem (Houston, TX, USA); nerve growth factor-7S (NGF) from Sigma-Aldrich (Burlington, MA, USA); rabbit polyclonal anti-TNKS1/2 antibody (H-350), mouse monoclonal anti-MAP2 antibody (AP20), mouse monoclonal anti-PAR antibody (10H), and ADP-HPD from Santa Cruz Biotechnology (Dallas, TX, USA); rabbit monoclonal anti-Axin1 antibody (C76H11), and rabbit monoclonal anti-unphosphorylated (Ser33/Thr41) β-catenin antibody (D13A1) from Cell Signaling Technology (Danvers, MA); DAPI, Alexa Fluor 488-conjugated goat anti-rabbit IgG, and Alexa Fluor 568-conjugated goat anti-mouse IgG from Thermo Fisher Scientific (Waltham, MA, USA)); and predesigned primers for real-time PCR from Takara Bio (Kusatsu, Japan).
Pnu 74654
Manufactured by Selleck Chemicals
Sourced in United States
PNU-74654 is a laboratory equipment product offered by Selleck Chemicals. It is designed for specific laboratory applications. The core function of PNU-74654 is to facilitate precise measurements and analysis within a controlled laboratory environment. Further details about the intended use or application of this product are not available.
Automatically generated - may contain errors
Lab products found in correlation
Mg132, by Selleck Chemicals (3 mentions)
Hanks balanced salt solution (hbss), by Boster Bio (2 mentions)
2 deoxy d glucose 2 dg, by Selleck Chemicals (2 mentions)
Chloroquine cq, by Merck Group (2 mentions)
3 methyladenine 3 ma, by Selleck Chemicals (2 mentions)
Umuc3, by American Type Culture Collection (2 mentions)
β catenin, by Santa Cruz Biotechnology (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Xav939, by Cayman Chemical (1 mentions)
Alexa fluor 568 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Abt888, by Apexbio (1 mentions)
Nerve growth factor 7s ngf, by Merck Group (1 mentions)
Icg 001, by Selleck Chemicals (1 mentions)
Alexa fluor 488 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Um uc 3, by Thermo Fisher Scientific (1 mentions)
Capmatinib, by Selleck Chemicals (1 mentions)
Icrt3, by Selleck Chemicals (1 mentions)
Trametinib, by Selleck Chemicals (1 mentions)
Xav939, by Selleck Chemicals (1 mentions)
10 protocols using pnu 74654
1
Examining Wnt Signaling Pathway
2
Evaluation of Autophagy Inhibitors on BLCA Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Two strains of BLCA cells (T24 and UM-UC-3) were obtained from American Type Culture Collection (Manassas, VA). Normally treated groups were grown in Roswell Park Memorial Institute (RPMI) 1640 medium for T24 cells or Dulbecco’s modified Eagle’s medium (DMEM) for UM-UC-3 cells supplemented with 10% foetal bovine serum (Gibco, Thermo Fisher Scientific, MA). Starvation-incubated groups were treated with Hank’s balanced salt solution (HBSS; Boster Biotechnology, Wuhan, China) for six hours and were then recovered in complete medium for further experiments [17 (link)]. Chloroquine (CQ, 20 mM; Sigma-Aldrich, USA), 3‐methyladenine (3-MA, 5 mM; Selleck Chemicals, Houston, TX), 2-deoxy-D-glucose (2-DG, 5 mM; Selleck Chemicals), MG132 (50 mM; Selleck Chemicals) and PNU-74654 (50 mM; Selleck Chemicals) were used to treat cell lines with different inhibitors.
3
Compound Screening in Drosophila Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Drugs and compounds were used as follows: trametinib (Selleckchem or biorbyt), QNZ (EVP4593), iCRT3, IWP-01, XAV-939, Capmatinib, JSH-23, PNU-74654 and Propidium iodide (PI) purchased from Selleckchem. Drug and compound stocks were diluted in DMSO or water; drugs were then mixed into standard fly food with final DMSO concentration 0.1% to prevent toxicity.
4
Evaluating Cytotoxic Compounds in Cells
5
Cell Culture and Genetic Manipulation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SW620, 293T, HCoEpiC, and RKO cells were cultured in DMEM containing 10% FBS (MilliporeSigma) and 1% penicillin/streptomycin. F-12K Nutrient Mixture (Invitrogen) was used to cultivate LoVo cells with 10% FBS and 1% penicillin/streptomycin. All cells were purchased from American Type Culture Collection and were detected by the mycoplasma detecting kit (Yise Medical Technology) to confirm a nonmycoplasma condition. All cells were cultured in the condition of 37°C and 5% CO2. The small interfering RNAs used in this study were siDicer-1, siDicer-2, and siDicer-3, which were purchased from RiboBio. The lentivirus-based shSURC-575, shSURC-1083, and shAPC (GenePharma) were used to infect SW620 and LoVo cell lines. The overexpression plasmid of SURC and APC were designed and purchased from GeneCopoeia. The miR–185-5p inhibitor and miR–185-5p mimic were purchased from RiboBio. Actinomycin D was purchased from MedChemExpress, while PNU-74654 and SKL2001 were purchased from Selleck.
6
Colorectal Cancer Cell Lines and Reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies used in this study were as following: β-catenin and caspase3 were purchased from Santa Cruz; GAPDH, cleaved caspase3, Axin1, Dvl2, GSK3β; TCF4 from Cell Signaling Technology and E-cadherin from BD Biosciences. XAV939 and VH-298 was purchased from MCE, MSAB from Sigma, PRI-724 and PNU-74654 from Selleck. Peptides used in this study, including SAHPA1, SAHPA1-VHLL, xStAx and xStAx-VHLL, were dissolved in DMSO for 10 mM stock solution.
Colorectal cancer cell lines SW480, HCT116, and LoVo were obtained from ATCC. SW480 cells were cultured in L15 medium supplemented with 10% FBS (Hyclone) in a 37 °C humidified incubator without CO2. HCT116 cells were cultured in McCoy’s 5 A medium supplemented with 10% FBS (Hyclone) in a 37 °C humidified incubator containing 5% CO2. LoVo cells were cultured in DMEM medium supplemented with 10% FBS (Hyclone) in a 37 °C humidified incubator containing 5% CO2.
Colorectal cancer cell lines SW480, HCT116, and LoVo were obtained from ATCC. SW480 cells were cultured in L15 medium supplemented with 10% FBS (Hyclone) in a 37 °C humidified incubator without CO2. HCT116 cells were cultured in McCoy’s 5 A medium supplemented with 10% FBS (Hyclone) in a 37 °C humidified incubator containing 5% CO2. LoVo cells were cultured in DMEM medium supplemented with 10% FBS (Hyclone) in a 37 °C humidified incubator containing 5% CO2.
7
Wnt/β-Catenin Signaling Modulation in JB6 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The JB6 cell line was used. At 24 h after seeding, PNU-74654 (Selleck, China), LiCl (Sigma, USA) or DMSO (Sigma, USA) was added into the culture medium. At 48 h after seeding, the cell pellets were collected following treatment with 0.25% trypsin. The total protein from the cell pellets were extracted respectively. SDS-PAGE was performed with 0.8% polyacrylamide gels. Proteins were then transferred to PVDF membranes (Millipore, USA), and membranes were blocked with 5% nonfat dry milk, in TBST (Tris-buffered saline containing 0.1% Tween-20). The PVDF membranes were then incubated with diluted primary antibodies overnight at 4 °C, washed with TBST, incubated with the HRP-labeled secondary antibodies (Sangon Biotech, China), and washed with TBST again. Finally, ECL (Bio-Rad, USA) was added to the PVDF membrane. The results were observed and recorded in a gel imager (Bio-Rad, USA). Primary antibodies against the following proteins were used: CyclinD1 (1:500, Sangon Biotech, China), β-Catenin (1:1000, Santa Cruz, USA), p-β-Catenin (1:5000, CST, USA), LEF1 (1:1000, Santa Cruz, USA). Anti-Smad 5 (1:1000, proteintech, China) was used to detect the expression of Smad5 protein after treatments.
8
Modulating Wnt Signaling in Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Wnt production inhibitor, IWP-2 (Selleck Chemicals), was used to inactivate Wnt signaling in NMR fibroblasts. NMR fibroblasts were plated at 500,000 cells/well in six-well plates. After 48 h, cells were treated with vehicle (0.1% DMSO) or IWP-2 in a dose-dependent manner (up to 100 μM), followed by lysate collection after 48 h. Lysates were then used for immunoblotting. To inhibit the interaction between β-catenin and TCF-4, NMR and MSFs were seeded for 48 h and then exposed to the small-molecule compound PNU-74654 (Selleck Chemicals) at concentrations of 50, 100, and 200 μM.
9
Investigating Akt and β-Catenin Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C-peptide was obtained from Peptron (Daejeon, Republic of Korea). The following reagents were used in this study: Cell tracking dye-kit-Green-Cytopainter (ab138891) from Abcam (Cambridge, United Kingdom); Vectashield Antifade Mounting Medium with 4,6-diamidino-2-phenylindole (DAPI) (H1200) from VECTOR (Malven, PA, United States); okadaic acid (OA) (ALX-350-003-C100) from Cell Signaling Technology (Danvers, MA, United States); 8-bromo adenosine 3ʹ5ʹ-cyclic adenosine monophosphate (8-Br-cAMP) (B5386) and Akti (124018) from Sigma-Aldrich (St. Louis, MO, United States); PNU-74654 (74654) from Selleckchem (Houston, Texas, United States); and MMP2/MMP9 inhibitor (ab1415190) from Abcam. Antibodies against Akt (9272), pS473-Akt (4051), β-catenin (9582), pS522-β-catenin (9566), pS33/37/T41-β-catenin (9561), pT202/Y204-ERK1/2 (9101), ERK1/2 (9102), MMP2 (87809), MMP9 (13667), and PP2A (2259) were obtained from Cell Signaling Technology. Antibodies against tubulin (ab11304) and PP1A (MAB3000) were purchased from Abcam and R&D Systems (Minneapolis, MN, United States), respectively. Anti-mouse secondary antibody (115-035-003) and anti-rabbit antibody (211-002-171) were obtained from Jackson ImmunoResearch Laboratories Inc. (West Grove, PA, United States).
10
Inhibition of Autophagy in BC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Two strains of BC cells (T24 and UM-UC-3) were obtained from American Type Culture Collection (Manassas, VA). Normal treated groups were grown in complete medium supplemented with 10% fetal bovine serum (Gibco, Thermo Fisher Scienti c, MA). Starvation incubated groups were treated by Hank's balanced salt solution (HBSS; Boster Biotechnology, Wuhan, China) for six hours and then recovered in complete medium for further experiments. Chloroquine (CQ, 20 mM; Sigma-aldrich, USA), 3methyladenine (3-MA, 5 mM; Selleck Chemicals, Houston, TX), 2-Deoxy-D-glucose (2-DG, 5 mM; Selleck Chemicals), MG132 (50 mM; Selleck Chemicals) and PNU-74654 (50 mM; Selleck Chemicals) were used to treat cell lines for different inhibitions.
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!