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Fh535

Manufactured by Merck Group
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Sourced in United States, Germany

The FH535 is a lab equipment product manufactured by Merck Group. It is a high-precision instrument designed for measuring and analyzing various physical and chemical properties of samples. The core function of the FH535 is to provide accurate and reliable measurement data for research, development, and quality control applications.

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Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions) Reporter plasmid, by Promega (2 mentions) Penicillin, by Thermo Fisher Scientific (2 mentions) Streptomycin, by Thermo Fisher Scientific (2 mentions) Cx 4945, by Merck Group (2 mentions) Hek293 cell, by American Type Culture Collection (2 mentions) Penicillin streptomycin, by Euroclone (2 mentions) Lithium chloride (licl), by Merck Group (2 mentions) Methyl 3h thymidine, by MP Biomedicals (2 mentions) Plko 1 puro vector, by Merck Group (2 mentions) Icg 001, by Selleck Chemicals (2 mentions) Hek293t, by American Type Culture Collection (2 mentions) Fetal bovine serum (fbs), by Merck Group (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Lumina 2, by PerkinElmer (1 mentions) Norcantharidin, by Merck Group (1 mentions) Sp600125, by Enzo Life Sciences (1 mentions) Cantharidin, by Enzo Life Sciences (1 mentions) Panc 1, by American Type Culture Collection (1 mentions) Bxpc 3, by American Type Culture Collection (1 mentions)

30 protocols using fh535

1

Xenograft Tumor Model in Nude Mice

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Four-week-old female BALB/c athymic nude mice were purchased from Shanghai SLAC Laboratory Animal Co. Ltd (Shanghai, China) and received humane care according to the Soochow University Institutional Animal Care and Treatment Committee. PANC-1 cells stably expressing firefly luciferase were injected into the left flanks of the mice in a total volume of 100 μL (0.5 × 107 cells), and the mice were randomly assigned to a dimethyl sulfoxide (DMSO) [intraperitoneally injected with 100 μL DMSO/DMEM (1:1)] or FH535 group [intraperitoneally injected with 25 mg/kg FH535 (Millipore) dissolved in 100 μL DMSO/DMEM (1:1)]. Treatment was conducted every 2 days for 20 days; tumor volume was measured with a caliper using the formula: volume = length × width2/2. At the end of the experiment, the mice were anaesthetized and given D-luciferin in PBS. Twenty minutes after the injection, bioluminescence was imaged with a charge-coupled device camera (IVIS; Lumina II, PerkinElmer). Then, the tumor tissue was stripped and formalin-fixed, paraffin-embedded, cut into 4-μm sections, and immunohistochemically stained.
Liu L., Zhi Q., Shen M., Gong F.R., Zhou B.P., Lian L., Shen B., Chen K., Duan W., Wu M.Y., Tao M, & Li W. (2016). FH535, a β-catenin pathway inhibitor, represses pancreatic cancer xenograft growth and angiogenesis. Oncotarget, 7(30), 47145-47162.
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2

Cantharidin-based Molecular Regulation

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Cantharidin, OA and SP600125 were purchased from Enzo Life Sciences International (Plymouth Meeting, PA, USA). NorCantharidin (NCTD) was purchased from Sigma (St. Louis, MO, USA). FH535 was purchased from Millipore (Billerica, MA, USA).
WU M.Y., XIE X., XU Z.K., XIE L., CHEN Z., SHOU L.M., GONG F.R., XIE Y.F., LI W, & TAO M. (2014). PP2A inhibitors suppress migration and growth of PANC-1 pancreatic cancer cells through inhibition on the Wnt/β-catenin pathway by phosphorylation and degradation of β-catenin. Oncology Reports, 32(2), 513-522.
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3

Culturing Pancreatic Cancer Cell Lines

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The human pancreatic cancer cell lines PANC-1 and BxPC-3 were purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA). The cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal calf serum (FCS), 100 U/mL penicillin, and 100 μg/mL streptomycin (Thermo Fisher Scientific) at 37°C in a 5% CO2 incubator under a humidified atmosphere; the cells were passaged every 2–3 days for exponential growth. FH535 was purchased from EMD Millipore (Billerica, MA, USA).
Wu M.Y., Liang R.R., Chen K., Shen M., Tian Y.L., Li D.M., Duan W.M., Gui Q., Gong F.R., Lian L., Li W, & Tao M. (2015). FH535 inhibited metastasis and growth of pancreatic cancer cells. OncoTargets and therapy, 8, 1651-1670.
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4

Analyzing Wnt Signaling in VSMC

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Effects of Pi on Wnt signaling in VSMC were analyzed using a luminescent reporter plasmid assay. Briefly, VSMC were transiently transfected with reporter plasmids (Promega, Mannheim, Germany) for the T-cell factor (TCF)/lymphoid enhancer-binding factor (LEF) binding motif (pGL4.49[luc2P/TCF-LEF RE/Hygro]) and a renilla vector (pGL4.74[hRluc/TK]). To inhibit Wnt activation, the β-Catenin/TCF site-inhibitor FH535 (10 µM; Merck) was used.
Freise C., Biskup K., Blanchard V., Schnorr J, & Taupitz M. (2024). Inorganic Phosphate-Induced Extracellular Vesicles from Vascular Smooth Muscle Cells Contain Elevated Levels of Hyaluronic Acid, Which Enhance Their Interaction with Very Small Superparamagnetic Iron Oxide Particles. International Journal of Molecular Sciences, 25(5), 2571.
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5

Investigating Wnt, EGF, and TGF-β Signaling

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For in vitro studies, stock solutions of MLN8237 (20 mmol/L), FH535 (10 mmol/L) and LY294002 (20 mmol/L) were prepared in DMSO. Stock solutions of recombinant human Wnt3a (100 μg/ml), EGF (100 μg/ml) and TGF-β1(100 μg/ml) were diluted in phosphate-buffered saline (PBS).
Dimethyl sulfoxide (DMSO), LY294002 and EGF were purchased from Sigma. FH535 was purchased from Merck. Wnt3a was purchased from abnova. MLN8237 was purchased from selleck.
AURKA, β-catenin, AKT1, p-AKT1, GSK-3β, p- GSK-3β, Twist, H3K4 me3/AC and H3K27 me2/me3/AC antibodies were purchased from Abcam. E-cadherin, N-cadherin, H3K4 me1/me2 and H3K27 me1 antibodies were purchased from Cell Signaling Technology. The Ki67 antibody was purchased from Santa Cruz Biotechnology. The H3 antibody was purchased from Ray Antibody. The GAPDH antibody was purchased from Zhongshan Bio Corp.
Liu X., Li Z., Song Y., Wang R., Han L., Wang Q., Jiang K., Kang C, & Zhang Q. (2016). AURKA induces EMT by regulating histone modification through Wnt/β-catenin and PI3K/Akt signaling pathway in gastric cancer. Oncotarget, 7(22), 33152-33164.
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6

Measuring MMP-2 and MMP-9 Activities

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Treatment dependent enzymatic activities of MMP-2 and MMP-9 in VSMC supernatants were measured by cleavage of 0.01 mg/ml dye-quenched DQ-gelatin (Molecular Probes, Life Technologies GmbH, Darmstadt, Germany) as described in detail earlier [18 (link)].
To determine effects of the Wnt inhibitor on enzymatic activities of MMP-2 and MMP-9, recombinant MMP-2 and MMP-9 alone or mixed with the Wnt inhibitor FH535 (10 μM; CAS 108409-83-2; Merck, Schwalbach, Germany) were subjected to DQ-gelatin measurements.
Freise C., Kretzschmar N, & Querfeld U. (2016). Wnt signaling contributes to vascular calcification by induction of matrix metalloproteinases. BMC Cardiovascular Disorders, 16, 185.
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7

Wnt Signaling Pathway Modulation in VSMCs

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VSMC were transiently transfected with reporter plasmids (Promega, Mannheim, Germany) for the T-cell factor (TCF)/lymphoid enhancer-binding factor (LEF) binding motif (pGL4.49[luc2P/TCF-LEF RE/Hygro]) and a renilla vector (pGL4.74[hRluc/TK]) as described [18 (link)]. As a positive control for Wnt activation the Wnt agonist I (CAS 853220-52-7; Merck, Schwalbach, Germany) was used. To inhibit Wnt activation, the β-Catenin/TCF site-inhibitor FH535 (10 μM; Merck) was used.
Freise C., Kretzschmar N, & Querfeld U. (2016). Wnt signaling contributes to vascular calcification by induction of matrix metalloproteinases. BMC Cardiovascular Disorders, 16, 185.
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8

Characterization of Medulloblastoma Cell Lines

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The MB cell lines Daoy (from American Type Culture Collection, Manassas, VA, USA) and Med1-MB (gift from Dr. Gerald Grant) were authenticated and tested for mycoplasma before experiments were conducted. Daoy and Med1-MB were cultured in Minimum Essential Media (MEM) supplemented with 10% fetal bovine serum (FBS) and 2 mM L-glutamine. HEK293 cells (from American Type Culture Collection, Manassas, VA, USA) were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum and 2 mM L-glutamine. FH-535, TBB, TBBz, and CX-4945 (Sigma, St. Louis, MO, USA) were used in inhibitor studies at stated concentrations for the specified time.
Nitta R.T., Bolin S., Luo E., Solow-Codero D.E., Samghabadi P., Purzner T., Aujla P.S., Nwagbo G., Cho Y.J, & Li G. (2019). Casein Kinase 2 inhibition sensitizes medulloblastoma to temozolomide. Oncogene, 38(42), 6867-6879.
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9

Cell Line Maintenance and Compound Treatments

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SW480 and SW620 cell lines were kindly gifted by Prof Alberto Bardelli, (University of Turin). HCT116 and HEK293T cells were purchased from ATCC. All cell lines were grown in DMEM supplemented with 10% FBS (Sigma), 2 mM L-glutamine and penicillin/streptomycin (10 U/ml) (Euroclone) at 37°C under a 5% CO2 atmosphere. FH535 (Sigma) and Cyclohexymide (Sigma) were dissolved in DMSO and diluted in colture medium at a concentration of 20 μM. and 25 μM respectively. In both cases, control cells received the same amount of DMSO.
Montorsi L., Guizzetti F., Alecci C., Caporali A., Martello A., Atene C.G., Parenti S., Pizzini S., Zanovello P., Bortoluzzi S., Ferrari S., Grande A, & Zanocco-Marani T. (2016). Loss of zfp36 expression in colorectal cancer correlates to wnt/ β-catenin activity and enhances epithelial-to-mesenchymal transition through upregulation of zeb1, sox9 and macc1. Oncotarget, 7(37), 59144-59157.
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10

Small Molecule Inhibitor Screening

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FH535, LiCl and BIO (6-bromoindirubin-3′-oxime) were purchased from Sigma-Aldrich. DKK1 was purchased from R & D systems.
Chang Y.H., Chu T.Y, & Ding D.C. (2017). WNT/β-Catenin signaling pathway regulates non-tumorigenesis of human embryonic stem cells co-cultured with human umbilical cord mesenchymal stem cells. Scientific Reports, 7, 41913.
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