Phtpp

Manufactured by MedChemExpress

Sourced in China

PHTPP is a laboratory equipment product provided by MedChemExpress. It is a chemical compound used as a tool in scientific research and experimentation.

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Lab products found in correlation

8 protocols using phtpp

Myoblast and Stem Cell Differentiation

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C2C12 myoblasts were seeded on culture plates in PM to achieve a 90% confluence, then the medium was changed to the basal differentiation medium (DM), supplemented with 20 µM epicatechin (EC, Sigma-Aldrich, Burlington, MA, USA), 10 µM genistein (GEN, Sigma-Aldrich), 10 µM luteolin (LUT, Sigma-Aldrich), 50 µM quercetin (QUE, Sigma-Aldrich), or 10 µM naringenin (NAR, Sigma-Aldrich) to induce differentiation. DM was DMEM containing 2% horse serum (HS; Thermo Scientific, Waltham, MA, USA) and 1% P/S. The differentiation lasted for 3 days, and the medium was changed every day.
PSCs were seeded on Matrigel-coated plates and cultured in PM for 2–3 days. When cells reached 90% confluence, differentiation was induced in DM supplemented with NAR at the indicated concentration for 5–7 days. In some experiments, ERβ antagonist PHTPP (MedChemExpress, Shanghai, China) at 10 μM was added to DM alone or in combination with NAR. The medium was changed every day.

Yan Q., Fei Z., Li M., Zhou J., Du G, & Guan X. (2022). Naringenin Promotes Myotube Formation and Maturation for Cultured Meat Production. Foods, 11(23), 3755.

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Ion Channel Modulation Reagents

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PPT, DPN, MPP, PHTPP, E₂, forskolin, genistein, rottlerin, wortmannin, GSK-7975A, ML-9, SKF-96365, CPA, and BAPTA-AM were purchased from MedChemExpress and dissolved in DMSO. CCh and indomethacin were purchased from Sigma-Aldrich. CCh was dissolved in ultrapure water. indomethacin was dissolved in anhydrous alcohol. All salts were supplied by Sangon Biotech and dissolved in ultrapure water.

Wan H., Li J., Chen X., Sellers Z.M, & Dong H. (2023). Divergent roles of estrogen receptor subtypes in regulating estrogen-modulated colonic ion transports and epithelial repair. The Journal of Biological Chemistry, 299(8), 105068.

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Cell Cycle Analysis of Genistin and PHTPP

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A total of 4 × 10⁴ cells were seeded into a 6-well plate and treated with genistin (5, 10, and 20 μM) and PHTPP (20 μM) (MedChemExpress, Monmouth Junction, NJ) for 5 days. The cells were collected, washed with PBS twice and permeabilized with ice-cold 70% ethanol at 4 °C until use. According to the manufacturer's protocol, the cells were washed twice and then treated with 100 μL trypsin inhibitor/RNase buffer using the CycleTEST™ PLUS DNA Reagent Kit (BD Biosciences, San Jose, CA, USA). After that, the cells were incubated with 300 μL PI stain solution for 10 min. Finally, the ploidies of the samples were detected by a BD FACSCanto II flow cytometer (BD Biosciences, San Jose, CA, USA) and analyzed by FlowJo software.

Tang X., Liao R., Zhou L., Yi T., Ran M., Luo J., Huang F., Wu A., Mei Q., Wang L., Huang X, & Wu J. (2024). Genistin: A Novel Estrogen Analogue Targeting ERβ to Alleviate Thrombocytopenia. International Journal of Biological Sciences, 20(6), 2236-2260.

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Comprehensive Evaluation of Pharmacological Agents

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All salts were from Sangon Biotech in Shanghai. Acetylcholine, N^ω-nitro-L-arginine and indomethacin were offered by Sigma-Aldrich (St Louis, USA). NE was purchased from GRANDPHARMA (China) co. LTD. 17β-Estradiol, LPS, Propyl pyrazole triol (PPT), MPP hydrochloride (MPP), G-1, G-15, Diarylpropionitrile (DPN), PHTPP, ouabain, TRAM-34, apamin, U73122, U73343, 2-APB and SKA-31 were supplied by MedChemExpress (MCE; New Jersey, USA), SN-6 was provided by Tocris (Bristol, UK). LiCl was supplied by Macklin (China). The human phospholipase C Beta 1 Polyclonal antibody (Cat No. 26551-1-AP) and human ITPR1-specific Polyclonal antibody (Cat No. 19962-1-AP) were from proteintech. The commercial kits of serum ALT, AST, LD and BUN were from Neobioscience (Jiancheng Bioengineering Institute, Nanjing, China). The commercial kit of serum DAO was from Solarbio. The commercial kit of serum Cr was from Beijing Leagene Biotechnology co. LTD.

Zhang L., Wan H., Zhang M., Lu W., Xu F, & Dong H. (2023). Estrogen receptor subtype mediated anti-inflammation and vasorelaxation via genomic and nongenomic actions in septic mice. Frontiers in Endocrinology, 14, 1152634.

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Investigating Estrogen Receptor Regulation in POA

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Four weeks after treatment, 60 rats (n = 20 per group) were randomly selected and equally divided into ERα group and ERβ group. Five rats were randomly selected from the SHAM group and OVX + E group of ERα group, the antagonists of ERα (AZD9496, HY-12870, MedChemExpress) were injected into POA, and the agonists of ERα (Propyl pyrazole triol, HY-100689, MedChemExpress) were injected into POA of the five rats randomly selected from the OVX group. Five rats were randomly selected from SHAM group and OVX + E group of ERβ group to inject antagonists of ERβ (PHTPP, HY-103456, MedChemExpress) into POA, and five rats from OVX group were randomly selected to inject agonists of ERβ (Liquiritigenin, HY-N0377, MedChemExpress) into POA. Other rats were given the same volume of cosolvent (10% DMSO + 40% PEG300 + 5% Tween-80 + 45% saline, MedChemExpress) in POA. After 72 h of drug injection, the POA was removed according to the above method, and the mRNA and protein expression of Vglut2 and Vgat in POA were detected by Western blot and qRT-PCR, respectively. The details on the primary and secondary antibodies of vglut2 and vgat are shown in Supplementary Table 1.1, and the details on the primers of vglut2 and vgat are shown in Supplementary Table 1.2.

Sun Y., Wang H., Wang W., Lu J., Zhang J., Luo X., Luan L., Wang K., Jia J., Yan J, & Qin L. (2022). Glutamatergic and GABAergic neurons in the preoptic area of the hypothalamus play key roles in menopausal hot flashes. Frontiers in Aging Neuroscience, 14, 993955.

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Calycosin Cytotoxicity Evaluated by MTT

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Cell viability was determined by MTT (Sigma-Aldrich; Merck KGaA) assay. Cells were harvested using 0.25% trypsin and seeded into 96-well plates at a density of 3×10⁴ cells/well at 37°C for 24 h. Then, cells were treated with calycosin (at concentrations of 0, 25, 50 or 100 µM) at 37°C for 24, 48 and 72 h, or with 100 µM calycosin in the presence or absence of the ERβ inhibitor PHTPP (50 µM; MedChemExpress) at 37°C for 48 h. In total, 20 µl MTT (5 mg/ml) was added to cells for 4 h at 37°C. Following incubation, DMSO (100 µl) was added to dissolve the formazan crystals and shaken at room temperature for 10 min. Subsequently, cell viability was assessed by measuring the absorbance at 570 nm using a microplate reader (Thermo Fisher Scientific, Inc.). Proliferation rate (%) was calculated as follows: Optical density (OD) treatment group/OD control ×100%.

Tian W., Wang Z.W., Yuan B.M, & Bao Y.G. (2020). Calycosin induces apoptosis in osteosarcoma cell line via ERβ-mediated PI3K/Akt signaling pathways. Molecular Medicine Reports, 21(6), 2349-2356.

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Estrogen Receptor Pathway Modulation

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Apigenin and histamine were from Sigma-Aldrich (St. Louis, MO, USA). The antibodies against Bax (1:1000), PI3K (1:1000), Akt (1:1000), and mTOR (1:1000) were obtained from Abbkine Scientific. (Wuhan, China). Antibodies for ERα (1:500), ERβ (1:500), p-PI3K (1:1000) and p-Akt (1:5000) were purchased from Abcam (Burlingame, city, CA, USA). AZD9496 (CAS No.: 1639042-08-2), PHTPP (CAS No.: 805239-56-9), PPT (CAS No.: 263717-53-9) and DPN (CAS No.: 1428-67-7) were obtained from MedChemExpress (Shanghai, China). FBS (Fetal bovine serum) was from Hyclone (Logan, UT, USA). Dulbecco’s modified Eagle medium (DMEM) was purchased from Gibco (Grand Island, NY, USA).

Zhang E., Zhang Y., Fan Z., Cheng L., Han S, & Che H. (2020). Apigenin Inhibits Histamine-Induced Cervical Cancer Tumor Growth by Regulating Estrogen Receptor Expression. Molecules, 25(8), 1960.

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Platelet Differentiation Assay

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A total of 4 × 10⁴ cells were seeded into a 6-well plate and treated with genistin (5, 10, and 20 μM), PHTPP (20 μM) (MedChemExpress, Monmouth Junction, NJ), LY294002 (10 μM) (MedChemExpress, Monmouth Junction, NJ), or SCH772984 (10 μM) (MedChemExpress, Monmouth Junction, NJ) for 5 days. The cells were harvested and washed once by ice-cold PBS. 5 μL of FITC conjugated anti-CD41 and PE conjugated anti-CD42b (Biolegend, San Diego, CA) were added to the cells and incubated on ice for 20 min in the dark. The labeled cells were washed twice and resuspended in 500 μL of PBS, subsequently analyzed by the BD FACSCanto II flow cytometer (BD Biosciences, San Jose, CA).

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