Latrunculin b
Manufactured by Cayman Chemical
Sourced in United States
Latrunculin B is a small molecule that inhibits actin polymerization. It is a natural product isolated from the Red Sea sponge Negombata magnifica. Latrunculin B binds to actin monomers, preventing them from assembling into filaments.
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Lab products found in correlation
Jasplakinolide, by Cayman Chemical (4 mentions)
Cytochalasin d, by Cayman Chemical (4 mentions)
Blebbistatin, by Cayman Chemical (2 mentions)
Dasatinib, by Selleck Chemicals (2 mentions)
Fasudil, by Selleck Chemicals (2 mentions)
Mrtf b, by Santa Cruz Biotechnology (2 mentions)
P egfr, by Cell Signaling Technology (2 mentions)
Donkey anti rabbit 680, by LI COR (2 mentions)
Donkey anti mouse 800, by LI COR (2 mentions)
Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (2 mentions)
Donkey anti goat 488, by Thermo Fisher Scientific (2 mentions)
Antibodies against yap1, by Cell Signaling Technology (2 mentions)
Antibodies against mrtf a, by Santa Cruz Biotechnology (2 mentions)
Donkey anti goat680, by LI COR (2 mentions)
Erlotinib, by Cayman Chemical (2 mentions)
Vemurafenib, by Selleck Chemicals (2 mentions)
Y 27632, by Selleck Chemicals (2 mentions)
Alexa fluor 546 phalloidin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
15 protocols using latrunculin b
1
Antifungal Compound Treatments on Conidia
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Latrunculin B (Cayman, USA) is dissolved in DMSO at a concentration of 25 mg/ml. Conidia incubated on the coverslips with hydrophobic surface were treated with LatB (final concentration 0.1 μg/ml) for 30 min, while the controls were treated with 5% DMSO. Then samples were washed with distilled water. Cycloheximide (MedChemExpress, USA) was solved in distilled water and the germinated conidia were treated with a final concentration 10 μg/ml for 10 min. Then samples were washed with distilled water. Benomyl (Aladdin, Shanghai, China) was solved in 0.1% DMSO and added to germinated conidia with a final concentration 1μg/ml. Then the samples were washed with distilled water. EGA (Merck, USA) was solved in 5% DMSO and was applied to samples with concentration 5 μg/ml for 1 h.
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2
Inducible EWS-FLI1 Knockdown in A673/TR/shEF Cells
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The dox-inducible A673/TR/shEF cell line was previously described.40 (link) Knockdown of EWS-FLI1 was achieved by addition of 1 μg/ml dox to the medium (24–72 h). The SK-N-MC EwS cell line was kindly provided by J Biedler (Memorial Sloan-Kettering Cancer Centre, New York, NY, USA). Cell lines were authenticated by STR profiling and regularly tested for mycoplasma (Mykoalert detection kit; Lonza, Basel, Switzerland). For serum induction cells were starved overnight in 0.2% fetal bovine serum medium with subsequent serum stimulation for 60 min with 20% fetal bovine serum medium. Latrunculin B (Cayman Chemicals, Ann Arbor, MI, USA) was added in serum-free DMEM at 1 μM concentration to the overnight starved cells 30 min before serum induction. Vehicle controls were treated with dimethyl sulfoxide (<0.05%).
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.
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.
5
Cytoskeletal Modulator Compound Characterization
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Vemurafenib (#S1267), Y-27632 (#S1049), fasudil (#S1573), and dasatinib (#S1021) were purchased from Sellekchem, Houston, TX, USA. Latrunculin B (#10010631), cytochalasin D (#11330), and erlotinib (#10483) were purchased from Cayman Chemical, Ann Arbor, MI, USA. Rho Inhibitor I (#CT04-A) was purchased from Cyoskeleton Inc, Denver, CO, USA. CCG-222740 18 (link) was synthesized in the lab of Dr. Scott Larsen at the University of Michigan. All compounds were diluted in DMSO to a stock concentration of 10 mM. Compound stock solutions were frozen at −20 °C. Antibodies against YAP1 (#14074), MLC2 (#3672), pMLC2 (#3674), Sox10 (#89356), and pEGFR (#3777) were purchased from Cell Signaling, Danvers, MA, USA. Antibodies against MRTF-A (#sc21558), MRTF-B (#sc98989), and Actin (#sc1616) were purchased from Santa Cruz, Dallas, TX, USA. Donkey anti-Mouse800 (#926-32212), Donkey anti-Goat680 (#926-68074), and Donkey anti-Rabbit680 (#926-68073) immunoblotting secondary antibodies were purchased from LI-COR, Lincoln, NE, USA. Alexa Fluor goat anti-rabbit488 (#A11034) and donkey anti-goat488 (#A11055) were purchased from Invitrogen. Alexa Fluor546 Phalloidin (#A22263) was purchased from ThermoFisher.
6
Cytoskeletal Modulator Compound Characterization
7
Inhibitory Compounds for Calcium Signaling
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Genistein (100 µg/ml), dynasore (80 µM), and 2-APB (100 µM) were purchased from Sigma and diluted in dimethyl sulfoxide (DMSO) to the given concentrations (Table 1 ). Verapamil (10 µM) and the cell-permeable cytosolic Ca2+ chelator BAPTA-AM (50 µM) were obtained from Calbiochem and diluted in DMSO. (−)-Xestospongin C (1 µM), the PLC inhibitor U73122 (10 µM), thapsigargin (10 µM), and ionomycin (2 µM) were purchased from Tocris Biosciences (diluted in DMSO). Latrunculin B was obtained from Cayman Chemical and diluted in ethanol. The scramblase inhibitor R5421, ethanimidothioic acid N-[(N-butylthio-N-methylamino)-carbonyloxy]-methyl ester (100 µM), was obtained from Endotherm and dissolved in DMSO (45 (link)). The final concentration of DMSO added to the cells was equal to or less than 0.01% of the total volume of the medium in all cases. Toxicity assays were performed to ensure that inhibitors did not have toxic effect when used with the cells (Fig. 7D ).
8
Investigating Cell Signaling Pathways
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Latrunculin B was purchased from Cayman Chemicals (Ann Arbor, MI, USA) and Phalloidin-iFluor 488 reagent (Cytopainter) was purchased from Abcam (Cambridge, MA, USA). Antibodies specific for BEX1 and BEX4 were purchased from Abcam and those specific for p-FAK, E-cadherin, N-cadherin, COL5A1, vimentin, YAP, TAZ, and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), penicillin, streptomycin, and TRIzol were obtained from Thermo Fisher Scientific (Waltham, MA, USA). siRNAs specific for mRNA of BEX1, BEX4, YAP, and TAZ were purchased from Bioneer (Daejeon, Republic of Korea) (Table S2 ).
9
Molecular Signaling Pathway Analysis
10
Characterization of Aortic Smooth Muscle Cells
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