ZCL278 was purchased from Tocris and diluted in DMSO at 50 mM. ICAM-1 recombinant protein (#ADP4-050), and fibronectin isolated from human plasma (1918-FN-02M) were purchased from R&D Systems. The type I solution of rat collagen, Lucifer Yellow CH dilithium salt, Resveratrol, and Lithium Chloride were purchased from Sigma-Merck. The recombinant human CCL2/MCP-1 Protein was obtained from R&D Systems and Miltenyi Biotec.
Zcl278
Manufactured by Bio-Techne
Sourced in United Kingdom
ZCL278 is a laboratory instrument used for cell culture applications. It functions as an incubator, providing a controlled environment for cell growth and maintenance. The device regulates temperature, humidity, and CO2 levels to support optimal cell culture conditions.
Automatically generated - may contain errors
Lab products found in correlation
Nsc23766, by Bio-Techne (3 mentions)
Ly294002, by Merck Group (2 mentions)
Rapamycin, by Merck Group (2 mentions)
Nsc23766, by Santa Cruz Biotechnology (2 mentions)
Lithium chloride (licl), by Merck Group (1 mentions)
Resveratrol, by Merck Group (1 mentions)
1918 fn 02m, by R&D Systems (1 mentions)
Lucifer yellow ch dilithium salt, by Merck Group (1 mentions)
Epidermal growth factor (egf), by Merck Group (1 mentions)
Nh4cl, by Merck Group (1 mentions)
U0126, by Merck Group (1 mentions)
Afatinib, by Selleck Chemicals (1 mentions)
Ml141, by Bio-Techne (1 mentions)
Cytochalasin d, by Bio-Techne (1 mentions)
Casin, by Bio-Techne (1 mentions)
5 n n dimethyl amiloride hydrochloride dma, by Merck Group (1 mentions)
5 n ethyl n isopropyl amiloride eipa, by Cayman Chemical (1 mentions)
Wiskostatin, by Bio-Techne (1 mentions)
Ck869, by Bio-Techne (1 mentions)
Ck666, by Bio-Techne (1 mentions)
12 protocols using zcl278
1
Investigating Cell Adhesion Molecules
2
Inhibitors of Actin Dynamics in Cell Biology
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All ligands, chemical compounds, and inhibitors used in this manuscript were cell biological grade: EGF (Sigma-Aldrich, cat# E9644), NH4Cl (Sigma-Aldrich, cat# A0171), NSC23766 (TOCRIS, cat# 2161), Casin (TOCRIS, cat# 5050), ZCL278 (TOCRIS, cat# 4794), ML141 (TOCRIS, cat# 4266), PBP10 (Calbiochem, cat# 529625), Rapamycin (Sigma-Aldrich, cat# S-015), U0126 (Sigma-Aldrich, cat# 19–147), Quercetagetin (Calbiochem, cat# 551590), Afatinib (Selleckchem, cat# S1011), 5-(N,N-Dimethyl)-amiloride hydrochloride (DMA, Sigma-Aldrich, cat# A4562), 5-(N-ethyl-N-isopropyl)-amiloride (EIPA, Cayman Chemical Company, cat# 14406), Wiskostatin (TOCRIS, cat# 4434), 187-1, N-WASP inhibitor (TOCRIS, cat# 2067), CK869 (TOCRIS, cat# 4984), CK666 (TOCRIS, cat# 3950), Cytochalasin D (TOCRIS, cat# 1233), and LY294002 (Sigma-Aldrich, cat# L9908).
3
Evaluating Rho GTPase Inhibitors on Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rho family GTPase inhibitors ML141 (Sigma-Aldrich), NSC23766 (Santa Cruz Biotech), ZCL278 (Tocris Bioscience), and Rhosin (Merck Milipore) were used at 15 μM, 50 μM, and 30 μM in i293-GFP and i293-GFP-ST and at 30 μM, 75 μM, and 60 μM in MCC13 cells. The integrin inhibitor RGDS (Tocris Bioscience) was used at a range of concentrations (see Results) on both 293-derived cells and MCC13 cells. Cell toxicity was measured using an MTS-based CellTiter 96 AqueousOne solution proliferation assay (Promega), as previously described (81 (link)).
4
Investigating Cell Signaling and Inhibition
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were stimulated or treated with different inhibitors as indicated in the figures. As stimuli, the complement C5a (20 nm ; R&D) as well as purified S100A8/S100A8‐homodimer (250 ng mL−1), S100A8/S100A9‐heterodimer (250 ng mL−1), and the mutated S100A8/S100A9‐N70A‐heterodimer (250 ng mL−1) were used. For the inhibition of the Rho GTPases, RhoA, Rac, and Cdc42, the cells were treated with the specific GEF inhibitors Rhosin hydrochloride (30 µm ), W56 (250 µm ), and ZCL278 (50 µm ) for 30 min, respectively (Tocris). Cryptotanshinone was used to inhibit the phosphorylation of STAT3 at a concentration of 5 µm for 30 min (Sigma–Aldrich).
5
Modulation of Cell Signaling Pathways
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell cultures were treated with 100 nM of the selective 5-HT7R agonist LP-211 (gift of M. Leopoldo, University of Bari, Italy), 100 nM of the HT7R antagonist SB-269970 (Tocris, Milan, Italy; Hagan et al., 2000 (link)), or with a combination of these drugs. Roscovitine (Sigma-Aldrich), a Cdk5 inhibitor, was used at the final concentration of 20 μM. The mTOR inhibitors rapamycin (Sigma-Aldrich) and torin 1 (Tocris), were used at a final concentration of 20 and 250 nM, respectively. ZCL 278 (Tocris), a selective inhibitor of Cdc42, was used at a final concentration of 50 μ M. Cytochalasin D (Sigma -Aldrich) was used at a final concentration of 100 nM, while latrunculin and jasplakinolide (Molecular Probes, Milan, Italy) were used at a final concentration of 2 μ M. Cells were pretreated for 30 min with 10 μM of U0126, the ERK 1/2 inhibitor, as recommended by manufacturer (Cell Signaling, Milan, Italy). Drugs were added to cultures 72 h after cell plating and incubated for appropriate time.
6
Cdc42 Inhibition in Cell Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
7
Immortalized Human Keratinocyte Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Keratinocytes (courtesy of Leena Bruckner-Tuderman, Department of Dermatology, University Hospital Freiburg) isolated from human foreskin were transformed with the human papillomavirus oncogenes E6 and E7 for immortalization (Kaur and McDougall, 1988 (link)). Cells were kept in Keratinocyte Serum-Free Medium (KSFM; Invitrogen, Carlsbad, CA) supplemented with 0.3 ng/ml recombinant EGF and 30 ng/ml bovine pituitary extract and 100 U/ml penicillin/streptomycin (culture medium). The experiment medium contained an additional 20 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (Sigma-Aldrich, St. Louis, MO). For pH experiments, the medium was adjusted to a pH of 7.45 by 2.5 N NaOH or to 6.6 by 10% HCl. The inhibitors CK-666 (Sigma-Aldrich), NSC-23766 (Tocris, Bristol, UK), ZCL-278 (Tocris), PIK-90 (Selleck Chemicals, Houston, TX), AG-1478 (Cayman Chemicals, Ann Arbor, MI), Y-27632 (Merck-Millipore, Billerica, MA), (–)-blebbistatin (Sigma-Aldrich), suramin (Tocris), pertussis toxin (Enzo Scientific, Farmingdale, NY), gallein (Tocris), telenzepine (Tocris), PPADS (Tocris), and CK-689 (Merck-Millipore) were used at the indicated concentrations.
8
Isolation and Culture of Dental Pulp Mesenchymal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The apical half of the dental pulp mesenchyme of the mandibular first molar was dissected out from wild-type mice at P3.5. The tissue was minced, centrifuged and resuspended in α-minimum essential medium (MEM) with 20% fetal bovine serum (FBS), 2 mM L-glutamine, 55 μM β-mercaptoethanol, 100 U/ml penicillin and 100 μg/ml streptomycin (Life Science Technologies). After 5 days of cultivation, cells were digested and obtained by passing through a 40 um strainer (Corning). The cell suspension was seeded at 3.5×104/well in 12-well plate culture (Corning) and incubated for 48 h at 37°C with 5% O2 and 5% CO2. The medium was then changed using fresh α-MEM without FBS for 10 h. Serum-starved mesenchymal cells were then incubated for another 20 h with DMSO as control or 50 μM ZCL278 (Cdc42 inhibitor; Tocris, 4794). For detection of PHH3, cells were washed with PBS before fixation with 4% PFA in PBS at room temperature for 20 min. Antibody to PHH3 was used at 1:100 dilution. Alexa Fluor 647 was used for detection. Samples were counterstained with DAPI.
9
Antagomir Treatment and Cellular Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pharmacological inhibitors used in this study are summarized in Fig. S1 and Table S1. HLEKs or hTCEpi cells were plated in a six-well plate. After overnight incubation, the cells were exposed to ir-antago, antago-103, or antago-107 (or mixture of antago-103 and antago-107) with and without an inhibitor. For the sequential treatment schedule, cells were treated with ir-antago, antago-103, or antago-107 for 24 h for the generation of vacuoles. After 24 h, each inhibitor was applied in cells. Amiloride, EIPA, roscovitine, manumycin A, Ro-32-0432, FAK inhibitor 14, NSC23766, EUK134, 8-bromo-cAMP, VU0155069, Go6983, Rottlerin, IBMX, and propranolol hydrochloride were obtained from Santa Cruz Biotechnology, Inc.; O-tricyclo[5.2.1.0(2,6)]dec-9-yl dithiocarbonate potassium salt (D609), l -α-phosphatidic acid sodium salt, and chloroquine were obtained from Sigma-Aldrich. BafA1 was from Cayman. PP2 was from EMD Millipore. ZCL278 was from Tocris Bioscience. SB203580 was from Cell Signaling Technology. BI-D1870 was from Enzo Life Sciences. Z-VAD-FMK was from BD. For short-term treatment of BafA1, cells were pretreated with BafA1 for 1 h, then medium with BafA1 was removed and cells were incubated in fresh medium with antagomirs for 24 h.
10
Inhibitor Effects on Cell Spreading
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Inhibitor experiments were performed in serum-containing medium, and treatments began 5 minutes prior to plating (spreading) or 5 minutes prior to scratch-wounding. Small-molecule inhibitors against signaling proteins, CDC42 (ZCL 278, 55 μM in DMSO), ERK (FR 180204, 3 μM in DMSO), FAK (PF 573228, 40 nM in DMSO), PI3K (LY294002, 5 μM in DMSO), RAC (EHT 1864, 600 nM in DMSO), and RHOA (CCG 1423, 3 μM in DMSO) were obtained from Tocris Bioscience, Bristol, UK. In order to ensure specificity of the inhibitors, dose-range studies were conducted with 3 doses for each inhibitor. Concentrations were chosen based on the lowest concentration for significant effect on spreading on any matrix. All inhibitors were used at a concentration less than 5x of the reported In vitro ID50 for each molecule.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!