Pri 724

Manufactured by Selleck Chemicals

Sourced in United States, Germany

PRI-724 is a laboratory equipment product designed for general scientific and research applications. It serves as a precision tool for various laboratory procedures. The core function of PRI-724 is to provide accurate and reliable measurements or performance in a controlled laboratory environment.

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Lab products found in correlation

Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (2 mentions) Xav939, by Selleck Chemicals (2 mentions) Alectinib, by Selleck Chemicals (2 mentions) Osimertinib, by Selleck Chemicals (2 mentions) Cellinsight high content microscope, by Thermo Fisher Scientific (2 mentions) Qgp 1, by Japanese Collection of Research Bioresources Cell Bank (1 mentions) Nci h727, by American Type Culture Collection (1 mentions) Lgk 974, by Novartis (1 mentions) Actilyse, by Boehringer Ingelheim (1 mentions) Truseq stranded mrna library preparation kit, by Illumina (1 mentions) Reliaprep rna tissue miniprep system, by Promega (1 mentions) Novaseq 6000, by Illumina (1 mentions) Il2rgammanull mice, by Jackson ImmunoResearch (1 mentions) Mouse igg1, by Cell Signaling Technology (1 mentions) Kg 501, by Bio-Techne (1 mentions) N acetylcysteine, by Selleck Chemicals (1 mentions) Urolithin a, by Selleck Chemicals (1 mentions) Z vad fmk, by Selleck Chemicals (1 mentions) Mg132, by Selleck Chemicals (1 mentions)

14 protocols using pri 724

Culturing and Treating Neuroendocrine Tumor Cells

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Human pancreatic NET cell line BON1 [9 (link)] (kindly provided by R. Göke, University of Marburg, Marburg, Germany) and pancreatic islet tumor cell line QGP-1 [9 (link)] (originally obtained from the Japanese Collection of Research Bioresources Cell Bank, Osaka, Japan), were both maintained in Dulbecco’s modified Eagle’s medium/F12 (at a ratio of 1:1) supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin and 0.4% amphotericin B in a 37 °C humidified incubator with 5% CO₂. Human bronchopulmonary neuroendocrine NCI-H727 tumor cells [9 (link)] (originally obtained from ATCC, Manassas, VA, USA) and the human small intestinal NET cell line GOT1 [9 (link),39 (link)] (kindly provided by O. Nilsson, Sahlgrenska University Hospital, Göteborg, Sweden) were both cultured in Roswell Park Memorial Institute medium-1640 (RPMI-1640) supplemented with 10% FBS, 1% penicillin/streptomycin, and 0.4% amphotericin B in a 37 °C humidified incubator with 5% CO₂.
To treat NET cells with WNT974 (also named LGK974; Novartis, Basel, Switzerland) or PRI-724 (Selleckchem, Germany), the cell lines were first seeded into cell culture dishes and grown overnight prior to treatment with various doses of WNT974 (1–32 µM, dissolved in dimethyl sulfoxide (DMSO)) or PRI-724 (1–10 µM, dissolved in DMSO) for different periods of time according to the assays listed below.

Jin X.F., Spöttl G., Maurer J., Nölting S, & Auernhammer C.J. (2020). Inhibition of Wnt/β-Catenin Signaling in Neuroendocrine Tumors In Vitro: Antitumoral Effects. Cancers, 12(2), 345.

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Ischemic Stroke Treatment Protocol

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Rats were randomly divided into five groups: sham group rats were treated with the same surgical operation without filament insertion, and they were administered 1 mL 1% DMSO intraperitoneally with 1 mL saline intravenously; vehicle group rats were treated with MCAO and administered 1 mL 1% DMSO intraperitoneally with 1 mL saline intravenously; rtPA group rats were treated with MCAO and administered 1 mL rtPA (10 mg/kg, Actilyse, Boehringer-Ingelheim, Germany) inserted into the left femoral vein 4 h after MCAO; rtPA + EX-4 group rats were treated with MCAO and administered EX-4 (100 µg/kg, MC Express, USA) intravenously immediately after rtPA injection; and rtPA + EX-4 + PRI-724 group rats were treated with MCAO and administered EX-4 intravenously and PRI-724 (10 mg/kg, Selleck, USA), a selective inhibitor of the β-catenin, intraperitoneally after rtPA injection.

Liu C., Sun S., Xie J., Li H., Li T., Wu Q., Zhang Y., Bai X., Wang J., Wang X., Li Z, & Wang W. (2022). GLP-1R Agonist Exendin-4 Protects Against Hemorrhagic Transformation Induced by rtPA After Ischemic Stroke via the Wnt/β-Catenin Signaling Pathway. Molecular Neurobiology, 59(6), 3649-3664.

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Transcriptomic Analysis of FLC Organoids

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Floating aggregates of FLC cells, referred to as FLC organoids, were prepared and cultured as described previously^[²⁰^] from the xenografts established using the patient‐derived FLC tumor line transplanted in 3‐month‐old female NOD‐scid IL2Rgamma^null mice (Jackson Laboratory) and were incubated in Kubota's Media (PhoenixSongs Biologicals) supplemented with 5 μm PRI‐724 (β‐catenin inhibitor; Selleck Chemicals) or DMSO for 24, 48, or 96 h. The protocol was approved by the Institutional Animal Care and Use Committee at CCHMC. Total RNA was extracted from FLC organoids using the ReliaPrep RNA Tissue Miniprep System (Promega). Poly‐A selected RNA was reverse‐transcribed using the Illumina TruSeq stranded mRNA library preparation kit and sequenced using Illumina NovaSeq 6000 (paired‐end 100 bp). Raw reads were aligned with the reference genome (GRCh38) using HISAT2,^[²¹^] and transcript/gene abundance was determined using Kallisto with annotations provided by University of California Santa Cruz using default parameters settings.^[²²^] All detected transcripts were tested for differential expression using a moderated t test with a cutoff false discovery rate < 0.05. Fusion transcripts identified in each sample using FusionCatcher^[²³^] along with regular transcripts were used to build an index in Kallisto.^[²²^] Differential transcript expressions were assessed with the R package DESeq2.^[²⁴^]

Gulati R., Johnston M., Rivas M., Cast A., Kumbaji M., Hanlon M.A., Lee S., Zhou P., Lake C., Schepers E., Min K., Yoon J., Karns R., Reid L.M., Lopez‐Terrada D., Timchenko L., Parameswaran S., Weirauch M.T., Ranganathan S., Bondoc A., Geller J., Tiao G., Shin S, & Timchenko N. (2022). β‐catenin cancer–enhancing genomic regions axis is involved in the development of fibrolamellar hepatocellular carcinoma. Hepatology Communications, 6(10), 2950-2963.

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DIPG and pedHGG Cell Culture Protocols

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DIPG and pedHGG cell lines were cultured as previously described^{9 (link)}. SF188 cells were cultured in DMEM/F12. To provide various cellular phenotypes that represent the heterogeneity of DIPG tumor samples and may differentially respond to drug treatment, cells were grown as gliomaspheres in tumor stem cell medium (TSM) or under differentiation conditions in TSM supplemented with 10% FCS, as recommended in ref. ^{9 (link)}. Unless stated otherwise, cells were treated with 2.5 µM ICG-001 (Calbiochem, Darmstadt, Germany), 2.5 µM PRI-724 (Selleckchem, Munich, Germany), and 0.25 µM (+)/–JQ1 (Selleckchem) dissolved in DMSO.

Wiese M., Hamdan F.H., Kubiak K., Diederichs C., Gielen G.H., Nussbaumer G., Carcaboso A.M., Hulleman E., Johnsen S.A, & Kramm C.M. (2020). Combined treatment with CBP and BET inhibitors reverses inadvertent activation of detrimental super enhancer programs in DIPG cells. Cell Death & Disease, 11(8), 673.

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Modulation of Type I Interferon Signaling

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Anti–mouse α-IFNAR was purchased from Leinco and was used at concentrations ranging from 0.1 μg/mL to 10 μg/mL, as indicated (I-401, clone MAR1-5A3). Mouse isotype control antibody (mouse IgG1, Cell Signaling Technology, 5415) was used as a negative control for this antibody where applicable. Nec-1 was used to inhibit necroptosis; it was purchased from Selleckchem and used at 50 μM, unless otherwise specified (catalog S8037). Pan-caspase inhibitor Z-VAD-FMK was used at 10 μM to block apoptosis and was purchased from Selleckchem (catalog S7023). MG132 was used at 10 μM to inhibit proteasomal degradation of proteins and was purchased from Selleckchem (catalog S2619). Pharmacological inhibitors of the CREB/CBP/β-catenin system were purchased from Selleckchem (PRI-724, catalog S8262; KG-501, catalog S8409; and C646, catalog S7152) and Tocris Biosciences (666-15, catalog 5661). Purified mouse IFN-β was purchased from PBL Assay Science (catalog 12405-1). Urolithin A was purchased from Selleckchem (catalog S5312) and was used as an inducer of mitophagy. N-acetylcysteine was purchased from Selleckchem (catalog S1623) and used as an antioxidant.

Agelidis A., Turturice B.A., Suryawanshi R.K., Yadavalli T., Jaishankar D., Ames J., Hopkins J., Koujah L., Patil C.D., Hadigal S.R., Kyzar E.J., Campeau A., Wozniak J.M., Gonzalez D.J., Vlodavsky I., Li J.P., Perkins D.L., Finn P.W, & Shukla D. (2021). Disruption of innate defense responses by endoglycosidase HPSE promotes cell survival. JCI Insight, 6(7), e144255.

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Comprehensive Biochemical Assay Protocol

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Olaparib, rucaparib, WNT-C59, and PRI724 were obtained from Selleckchem. Pyrvinium pamoate was obtained from Sigma Aldrich. The following antibodies were obtained from the indicated suppliers: BRCA2 (Bethyl, Cat#A303-434A, 1:2000), Vinculin (Cell Signaling Technology, Cat#13901, 1:1000), WNT3A (R&D systems, Cat# MAB9025-100, 1:1000), γH2Ax (Ser139) (EMD Millipore, Cat# 05-636, 1:1000), mouse anti-β-Actin (Abcam, Cat# ab6276, 1:10,000), Rabbit anti-β-Actin (Abcam, Cat# ab8227, 1:10,000), Rad51 (Abcam, Cat# ab176458). Anti-rat HRP (Jackson ImmunoResearch, Cat# 112-035-062, 1:5000)

Yamamoto T.M., McMellen A., Watson Z.L., Aguilera J., Ferguson R., Nurmemmedov E., Thakar T., Moldovan G.L., Kim H., Cittelly D.M., Joglar A.M., Brennecke E.P., Wilson H., Behbakht K., Sikora M.J, & Bitler B.G. (2019). Activation of Wnt signaling promotes olaparib resistant ovarian cancer. Molecular carcinogenesis, 58(10), 1770-1782.

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Colorectal Cancer Cell Lines and Reagents

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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.

Liao H., Li X., Zhao L., Wang Y., Wang X., Wu Y., Zhou X., Fu W., Liu L., Hu H.G, & Chen Y.G. (2020). A PROTAC peptide induces durable β-catenin degradation and suppresses Wnt-dependent intestinal cancer. Cell Discovery, 6, 35.

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Evaluating Combination Therapies for Cancer

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Small molecule inhibitors were all purchased commercially from Selleck Chemicals, and included Osimertinib (S7297), Alectinib (S2762), XAV-939 (S1180), and PRI-724 (S8262). Dimethyl sulfoxide (DMSO) (Fisher Scientific) was used to dissolve small molecule inhibitors according to manufacturer’s recommendations for use in in vitro experiments. PC9 and H3122 cells (5 × 10³) were seeded in 96-well plate format (μclear CellStar, Greiner) and rested for 24 hours before treatment. Treatment included: i) DMSO, ii) tyrosine kinase inhibitors (TKI) Osimertinib (PC9 cells) or Alectinib (H3122 cells), iii) Wnt/β-catenin inhibitors PRI-724 or XAV-939, and v) indicated combination therapies of TKI and Wnt/ β-catenin inhibitors. All conditions were plated in technical quadruplicate and cells were retreated every 3 days. At each imaging interval, cellular nuclei were stained with Hoechst 33342 (Thermo Fisher Scientific) and scanned using a CellInsight High-Content Microscope (Version 6.4.3 Build 7204, Thermo Fisher Scientific) with a 4X objective.

Maynard A., McCoach C.E., Rotow J.K., Harris L., Haderk F., Kerr D.L., Yu E.A., Schenk E.L., Tan W., Zee A., Tan M., Gui P., Lea T., Wu W., Urisman A., Jones K., Sit R., Kolli P.K., Seeley E., Gesthalter Y., Le D.D., Yamauchi K.A., Naeger D.M., Bandyopadhyay S., Shah K., Cech L., Thomas N.J., Gupta A., Gonzalez M., Do H., Tan L., Bacaltos B., Gomez-Sjoberg R., Gubens M., Jahan T., Kratz J.R., Jablons D., Neff N., Doebele R.C., Weissman J., Blakely C.M., Darmanis S, & Bivona T.G. (2020). Therapy-Induced Evolution of Human Lung Cancer Revealed by Single-Cell RNA Sequencing. Cell, 182(5), 1232-1251.e22.

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Pharmacological Inhibitors for Cell Research

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Trametinib, ICG-001, PRI-724, dabrafenib, GDC-0994, PD318088, selumetinib were purchased from Selleckchem. Erlotinib, SCH772984 and RAF265 were purchased from Cayman Chemical. Gefitinib was purchased from BioVision. XAV939 and CHIR99021 were purchased from Merck Millipore.

Zhan T., Ambrosi G., Wandmacher A.M., Rauscher B., Betge J., Rindtorff N., Häussler R.S., Hinsenkamp I., Bamberg L., Hessling B., Müller-Decker K., Erdmann G., Burgermeister E., Ebert M.P, & Boutros M. (2019). MEK inhibitors activate Wnt signalling and induce stem cell plasticity in colorectal cancer. Nature Communications, 10, 2197.

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Investigating Combination Therapies for NSCLC

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Small molecule inhibitors were all purchased commercially from Selleck Chemicals, and included Osimertinib (S7297), Alectinib (S2762), XAV-939 (S1180), and PRI-724 (S8262). Dimethyl sulfoxide (DMSO) (Fisher Scientific) was used to dissolve small molecule inhibitors according to manufacturer’s recommendations for use in in vitro experiments. PC9 and H3122 cells (5 × 10³) were seeded in 96-well plate format (jclear CellStar, Greiner) and rested for 24 hours before treatment. Treatment included: i) DMSO, ii) tyrosine kinase inhibitors (TKI) Osimertinib (PC9 cells) or Alectinib (H3122 cells), iii) Wnt/β-catenin inhibitors PRI-724 or XAV-939, and v) indicated combination therapies of TKI and Wnt/ β-catenin inhibitors. All conditions were plated in technical quadruplicate and cells were retreated every 3 days. At each imaging interval, cellular nuclei were stained with Hoechst 33342 (Thermo Fisher Scientific) and scanned using a CellInsight High-Content Microscope (Version 6.4.3 Build 7204, Thermo Fisher Scientific) with a 4X objective.

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