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Jasplakinolide

Manufactured by Cayman Chemical
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Jasplakinolide is a chemical compound used as a research tool in cell biology. It functions as an actin-stabilizing agent, binding to and stabilizing actin filaments within cells.

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Lab products found in correlation

Latrunculin a, by Cayman Chemical (4 mentions) Latrunculin b, by Cayman Chemical (4 mentions) Blebbistatin, by Cayman Chemical (3 mentions) Cytochalasin d, by Cayman Chemical (3 mentions) Pf 573228, by Selleck Chemicals (2 mentions) Its g 100x, by Thermo Fisher Scientific (2 mentions) Ionomycin, by Thermo Fisher Scientific (2 mentions) Yohimbine, by Merck Group (2 mentions) Dasatinib, by Cayman Chemical (2 mentions) Anisomycin, by Merck Group (2 mentions) Gefitinib, by Cayman Chemical (2 mentions) Bapta am, by Selleck Chemicals (2 mentions) Ly2584702, by Selleck Chemicals (2 mentions) Pf562271, by Adipogen (2 mentions) Gdc 0994, by Apexbio (2 mentions) Zstk474, by Cell Signaling Technology (2 mentions) Uk14304, by Merck Group (2 mentions) Y 27632, by Enzo Life Sciences (2 mentions) Latrunculin b, by Enzo Life Sciences (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)

15 protocols using jasplakinolide

1

Topical Application of Pharmacological Agents

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The PF573228, blebbistatin, or jasplakinolide (Cayman, Ann Arbor, MI, USA) were dissolved in DMSO. Then, 20 μL was applied once per day directly to the wound surface, starting at PWD 10 and continuing until PWD 17.
Harn H.I., Chiu P.Y., Lin C.H., Chen H.Y., Lai Y.C., Yang F.S., Wu C.C., Tang M.J., Chuong C.M, & Hughes M.W. (2022). Topological Distribution of Wound Stiffness Modulates Wound-Induced Hair Follicle Neogenesis. Pharmaceutics, 14(9), 1926.
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2

Decellularized Extracellular Matrix Effects on Cardiac Explants

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Hearts harvested from day 1 mice were washed in cold 1 x PBS. Atrial was removed and ventricle was cut to approximately 1mm3 pieces using surgical scissors. Two microliters of adult or fetal dECM were injected into explants using 10μl Hamilton syringes with 23G needles before plating on 48-well plates coated with 1.5mg/ml collagen I (Corning) gel. Explants were incubated at 37°C for 2h to induce dECM polymerization and explant-collagen gel attachment. Explants were cultured in explant culture media [1% fetal bovine serum, 2mM L-glutamine, 1 x insulin-transferrin-selenium (Gibco, ITS-G 100X) in M199] for 6 days with 10μM BrdU labeling for 3 days before fixation. Paraformaldehyde (4%) was used for fixation. In separate experiments, explants were cultured in explant culture media supplemented with 5mM ribose or 0.2mM BAPN for 6 days from plating in combination with dECM treatment conditions. Atomic force microscopy was used to measure the elastic modulus of decellularized explants [13 (link)], [32 (link)].
In order to regulate cytoskeleton polymerization, explants were cultured in explant culture media supplemented with Jasplakinolide (Cayman), Latrunculin A (Cayman), Y27632 (Chemdea), PF573228 (Selleck) for 3 days; no BrdU labeling applied.
Wang X., Pierre V., Liu C., Senapati S., Park P.S, & Senyo S.E. (2021). Exogenous extracellular matrix proteins decrease cardiac fibroblast activation in stiffening microenvironment through CAPG. Journal of molecular and cellular cardiology, 159, 105-119.
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3

Fluorescent Microscopy Imaging of CD93

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Lympholyte-poly and Cy3-conjugated rabbit-anti-mouse Fab fragments were purchased from Cedarlane. Ham’s F12, DMEM, 100X antibiotic-antimycotic solution and fetal bovine serum (FBS) were purchased from Wisent. #1.5 thickness, 18 mm round coverslips were purchased from Electron Microscopy Supplies. Anti-human CD93 (clone R139) was purchased from eBioscience. Leiden chambers were purchased from Quorum instruments. Latrunculin B, methyl-β-cyclodextran, blebbistatin and jasplakinolide were purchased from Cayman chemical. Cholesterol was purchased from Advanti Polar Lipids. GenJet Plus In Vitro DNA Transfection Reagent was purchased from SignaGen Laboratories. The Leica DMI6000B microscope and all components were purchased from Leica Microsystems. Matlab equipped with the parallel processing, statistics, image processing, and optimization toolboxes was purchased from Mathworks. All other materials were purchased from Thermo Fisher.
Goiko M., de Bruyn J.R, & Heit B. (2016). Short-Lived Cages Restrict Protein Diffusion in the Plasma Membrane. Scientific Reports, 6, 34987.
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4

Actin Cytoskeleton Manipulation and Visualization

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0.5 × 106 J8-GECI cells were washed ×1 in PBS and resuspended in either 10 μM DMSO (control), 1 μM latrunculin B, and 10 μM cytochalasin D, or 100 nM jasplakinolide (Cayman Chemical) diluted in RPMI (no supplements) for 1 h at 37  °C with 5% CO2. Cells were then fixed in PBS containing 4% PFA and 0.25% glutaraldehyde for 20 min at room temperature. During this period the cells were also labeled with CellMask Deep Red Plasma Membrane Stain (ThermoFisher). Cells were washed ×2 in PBS and immediately placed on PLL-coated glass surfaces (previously washed with ethanol) for 10 min prior to imaging at room temperature. Images were taken as described for imaging of fixed T cell microvilli.
Jenkins E., Körbel M., O’Brien-Ball C., McColl J., Chen K.Y., Kotowski M., Humphrey J., Lippert A.H., Brouwer H., Santos A.M., Lee S.F., Davis S.J, & Klenerman D. (2023). Antigen discrimination by T cells relies on size-constrained microvillar contact. Nature Communications, 14, 1611.
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5

Jasplakinolide Effects on Glioma Spheroids

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500 DiD labeled SMA560 glioma cells were seeded in spheroid-forming conditions per well of a 96-well plate. 18 h prior to implantation, spheroids or brain slices were treated with 1 μM jasplakinolide (Cayman #11705) or DMSO (Sigma # 41639-100ML). 24 h after implantation, brain slices were fixed and imaged by confocal microscopy.
Cell viability in vitro was measured with trypan blue staining (Sigma # 93595-50ML).
Eisemann T., Costa B., Strelau J., Mittelbronn M., Angel P, & Peterziel H. (2018). An advanced glioma cell invasion assay based on organotypic brain slice cultures. BMC Cancer, 18, 103.
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6

Monitoring HeLa Cell Translation and Signaling

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Human HeLa-kyoto cells were obtained from ATCC and maintained in high glucose DMEM supplemented with 10% fetal bovine serum (Atlas). For translation inhibition, cells were treated at 50 μg/ml with puromycin (Sigma) for 20 min or as otherwise indicated. Cells for MKL nuclear-cytoplasmic assessment were treated with 2μM cytochalasin D (Cayman Chemical), 1μM Jasplakinolide (Cayman Chemical) or dimethyl sulfoxide (Sigma) for controls, for 2h prior to fixation for analyses. For serum stimulation experiments cells were starved for 18–24 h in medium containing 0.3% serum and stimulated by replacing with medium containing 15% serum.
Wiggan O, & Stasevich T.J. (2024). Single molecule imaging of the central dogma reveals myosin-2A gene expression is regulated by contextual translational buffering. bioRxiv.
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7

Chemical Compound Preparation Protocol

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Stocks of 200 μM phorbol-12,13-dibutyrate (PDBu, EMD Millipore, #524390), 1 mg/mL anisomycin (Sigma-Aldrich, A9789), 10 mM UK14304 (Sigma-Aldrich, U104), 10 mM yohimbine (Sigma-Aldrich, Y3125), 1 mM gefitinib (Cayman, #13166), 1 mM ionomycin (Peprotech, #5608212), 2 mM jasplakinolide (Cayman, #11705), 25 mM latrunculin B (Enzo Life Sciences, BML-T110–0001), 10 mM PF562271 (AdipoGen, SYN-1064), 10 mM ZSTK474 (Cell Signaling, #13213), 10 mM dasatinib (Cayman, #11498), 10 mM GDC-0994 (APExBIO Technology, B5817), 1 mM LY2584702 (Selleck, S7698), 10 mM BAPTA-AM (Selleck, S7534) were prepared by dissolving the chemicals in DMSO; 10 mM Y27632 (Enzo Life Sciences, #ALX-270–333) in water. Stocks were diluted to the indicated final concentrations in the culture medium. The EGF stock solution was prepared by dissolving EGF (Sigma-Aldrich, E9644) in 10 mM acetic acid to a final concentration of 1 mg/ml. All drug stocks were stored at −20°C. 2-Deoxyglucose (2-DG, MilliporeSigma, #D8375) was dissolved in culture medium to 100 mM and used immediately.
Yang J.M., Chi W.Y., Liang J., Takayanagi S., Iglesias P.A, & Huang C.H. (2021). Deciphering cell signaling networks with massively multiplexed biosensor barcoding. Cell, 184(25), 6193-6206.e14.
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8

Membrane Protein Regulation Assay

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All reagents were purchased from Sigma-Aldrich unless otherwise stated. Drugs were made up in DMEM (Life Technologies), and cells were pretreated at 37 °C, 5% CO2 in a humidified incubator. MCD and α-cyclodextrin (αCD) were used at 5 mm for 15 min, and water-soluble cholesterol was used at 100 μg/ml for 30 min. Pan-caspase inhibitor, zVAD-FMK, was purchased from BioVision Inc. (Milpitas, CA) and used at 5 μm for 1 h. Latrunculin-A was used at 5 μm for 30 min. Jasplakinolide was used at 30 nm for 1 h, and (−)-blebbistatin was used at 100 μm for 1 h; both were purchased from the Cayman Chemical Company (Ann Arbor, MI). For bromopalmitate (BrP), a stock solution of 100 mm was made up fresh in DMSO each time and diluted 1:1000 in DMEM plus serum to give a final concentration of 100 μm for 16 h. For carbenoxolone treatment, cells were not pretreated, but 30 μm was present in all solutions used in the experiment. Rabbit polyclonal P2X7 C terminus antibody was obtained from Alomone Labs (Jerusalem, Israel), and anti-mouse and anti-rabbit HRP-conjugated secondary antibodies were from Thermo Fisher Scientific and Bio-Rad, respectively.
Robinson L.E., Shridar M., Smith P, & Murrell-Lagnado R.D. (2014). Plasma Membrane Cholesterol as a Regulator of Human and Rodent P2X7 Receptor Activation and Sensitization. The Journal of Biological Chemistry, 289(46), 31983-31994.
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9

Molecular Signaling Pathway Analysis

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Cholesterol and methyl-β-cyclodextrin were purchased from Sigma-Aldrich. Compound C, STO-609 acetate and CEP-1347 were purchased from Tocris Bioscience. Cytochalasin D, Jasplakinolide and Latrunculin B were purchased from Cayman Chemical. PD98059 was purchased from Enzo Life Sciences. SP600125 and U0126 were purchased from Calbiochem. Antibodies for phospho-AMPKα (#2535), MyD88 (#4283), phospho-ERK1/2 (#9101), total-ERK1/2 (#9102), phospho-JNK1/2 (#9251), total-JNK1/2 (#9252), anti-rabbit HRP-linked antibody (#7074) and anti-mouse HRP-linked antibody (#7076) were purchased from Cell Signaling Technology. Antibodies for AMPKα1 (sc-19128), MUC5AC (sc-21701), TAK1 (sc-7967), MLK3 (sc-166592), c-myc (sc-40), β-actin (sc-8432) and donkey anti-goat HRP-linked antibody (sc-2020) were purchased from Santa Cruz Biotechnology. Donkey anti-rabbit IgG Alexa Fluor 488 (A-21206) and goat anti-mouse IgG Alexa Fluor 546 (A-11030) was purchased from Thermo Fisher Scientific. The human validated short interfering RNA (siRNA) oligonucleotides for AMPKα1 (M-005027-02-0005), TAK1 (M-003790-06-0005) and control siRNA (D-001206-14-05) were purchased from Dharmacon.
Matsuyama S., Komatsu K., Lee B.C., Tasaki Y., Miyata M., Xu H., Shuto T., Kai H, & Li J.D. (2022). Negative cross-talk between TLR2/4-independent AMPKα1 and TLR2/4-dependent JNK regulates S. pneumoniae-induced mucosal innate immune response. Journal of immunology (Baltimore, Md. : 1950), 209(8), 1532-1544.
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10

Preparation and Application of Pharmacological Agents

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Stock reagents were prepared according to the manufacturer’s recommendation in water, DMSO (<0.01% final volume during experiments), or phosphate buffered saline (PBS), stored at −20°C, and diluted into ACSF or intracellular recording solutions as needed. CNQX, D-APV, SR 141716, and WIN 55,212–2 were acquired from the NIMH Chemical Synthesis and Drug Supply Program; salts for making cutting, ACSF, ziram, and intracellular recording solutions from Sigma Aldrich (St. Louis, MO); AM251, NSC-23766, MG-132, DAMGO, cycloheximide from Tocris Bioscience (Bristol UK); jasplakinolide, anisomycin, PYR41 from Cayman Chemical (Ann Arbor, MI); CK-666 from EMD Millipore. Reagents were either acutely bath applied, diluted into the intracellular recording solution, or preincubated with slices/cultures, as indicated in Results.
Monday H.R., Bourdenx M., Jordan B.A, & Castillo P.E. (2020). CB1-receptor-mediated inhibitory LTD triggers presynaptic remodeling via protein synthesis and ubiquitination. eLife, 9, e54812.
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