Primary antibody for GAPDH was from Bioworld (Minneapolis, MN), for LC3, p62, Vps34, Beclin-1 and Bcl-2 from Cell signaling (Beverly, MA. Wortmannin (WM) was purchased from sigma Aldrich (St Louis, MO). The supersignal west pico chemiluminescent substrate was obtained from Pierce (Rockford, IL).
Wortmannin wm
Manufactured by Merck Group
Sourced in United States, Italy
Wortmannin (WM) is a fungal metabolite that has been widely used as a chemical inhibitor in research laboratories. It functions as a potent and selective inhibitor of various phosphoinositide 3-kinase (PI3K) enzymes, which play crucial roles in cellular signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
Ns 398, by Cayman Chemical (2 mentions)
Beclin 1, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Primary antibody for gapdh, by Bioworld Technology (1 mentions)
Vps34, by Cell Signaling Technology (1 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Lps escherichia coli 0111 b4, by Merck Group (1 mentions)
Verteporfin, by MedChemExpress (1 mentions)
Insulin like growth factor 1 igf 1, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Jsh 23, by Cayman Chemical (1 mentions)
Il 33, by Promega (1 mentions)
U0126, by Promega (1 mentions)
17β estradiol e2, by Merck Group (1 mentions)
G 15 3as 4r 9br 4 6 bromo 1 3 benzodioxol 5 yl 3a 4 5 9b 3h cyclopenta c quinolone, by Bio-Techne (1 mentions)
Sta 21, by Santa Cruz Biotechnology (1 mentions)
G 1 1 4 6 bromobenzol 1 3 diodo 5 yl 3a 4 5 9b tetrahidro3h5cyclopenta c quinolin 8yl ethanone, by Bio-Techne (1 mentions)
Cycloheximide (chx), by MedChemExpress (1 mentions)
Mg132, by MedChemExpress (1 mentions)
12 protocols using wortmannin wm
1
Investigating Autophagy Regulators in Cells
2
Endothelial Cell Inflammation Assay
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Escherichia coli LPS (0111:B4) and Wortmannin (WM) were purchased from Sigma-Aldrich (St. Louis, MO, USA). A Bicinchoninic Acid protein assay kit and supersignal west pico chemiluminescent substrate were obtained from Pierce (Rockford, IL, USA). MTT [3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] reagent was from Bio Besic Inc. (Markham, ON, Canada). Foetal bovine serum (FBS) was obtained from Gibco (Grand Island, NY, USA). Antibody information: against α-tubulin, from Sigma-Aldrich; against total Akt and phosphor-Akt (p-Akt), from Cell Signaling Technology (Beverly, MA, USA); against E-selectin, from Abcam (Cambridge, UK); against HSPA12B, ICAM-1, IL-6, TNF-α and VCAM-1, from Santa Cruz Biotechnology (Santa Cruz, CA, USA); against COX-2, from R&D Systems Inc (Minneapolis, MN, USA). ELISA kit was purchased from Yuanye Bio-Technology Co., Ltd (Shanghai, China).
3
Molecular Inhibitors of IGF-1 Signaling
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Insulin-like growth factor-1 (IGF-1) and PI3K inhibitor Wortmannin (WM) were purchased from Sigma-Aldrich (Milan, Italy). The dual insulin-like growth factor 1 receptor (IGF-1R)/insulin receptor (IR) inhibitor OSI-906 (Linsitinib) was obtained from Tocris Bioscience (Space, Milan, Italy). Focal Adhesion Kinase selective inhibitor VS-4718 (PND-1186) was bought from Santa Cruz Biotechnology (DBA, Milan, Italy). The YAP/TEAD complex suppressor Verteporfin was purchased from Med Chem Express (DBA, Milan, Italy). All the aforementioned compounds were dissolved in dimethyl-sulfoxide (DMSO) except IGF-1 which was dissolved in water.
4
Cellular DNA Damage Response
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Neocarzinostatin (NCS was a generous gift of V. Favaudon (Institut Curie, Orsay, France). NU7026 and KU55933 (Calbiochem, France) and wortmannin (WM, Sigma, France) were prepared in dry dimethyl sulfoxide and stored at −20°C. Cells were pre-treated with kinase inhibitors for 18 hours and either γ-irradiated at 10 Gy or treated with NCS.
5
Pharmacological Inhibition of Signaling Pathways
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NS398 (COX-2 inhibitor) was purchased from Cayman Chemical (Ann Arbor, MI, USA). ONO-AE3-208 (selective EP4 antagonist, EP4A) was a gift from ONO Pharmaceuticals, Osaka, Japan. Wortmannin (WM), an irreversible PI3K inhibitor purchased from Sigma-Aldrich (Saint Louis, MO, USA), Dr. Lala generously shared 1 mM of each drug with us. For all treatments in vitro, DMSO (vehicle) served as the control.
6
Luminol-based ROS Detection in Plants
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Luminol-based ROS production in leaf tissue was performed as described [38] (link) using indicated PAMP concentrations or using Pto DC3000 or Pto hrcC−[42] (link). For inhibitor treatments, leaf tissue was pre-treated by floating leaf discs in water-containing 30 µM Wortmannin (Wm; Sigma-Aldrich; St. Louis, MO) for one hour prior to subsequent flg22-elicitation [13] (link). For ROS assays in six-to-seven-day old plants, cotyledons were cut in half, and the two halves were placed into a 96-well microplate for elicitation. All ROS experiments shown in a same panel were performed in the same 96-well plate at the same time to allow for direct comparison.
7
IL-33 Modulates UVB-Induced NF-κB Signaling
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PAM212 cells (Lonza Walkersville, Inc.) were seeded in a medium composed of Eagle's minimal essential media, 10% FBS, l-glutamine, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid buffer, sodium pyruvate, nonessential amino acids, and penicillin-streptomycin (Gibco, Waltham, MA, USA). First, cells were treated with 24 h of starvation in a 2% FBS-added medium. Then, UVB irradiation (5.5 mJ/cm2) was performed with the use of an UVB device (302 nm) supplied by Rayminder, Ottawa Hills, OH, USA. Following this, cells in the stimulation groups were given 100 ng/ml recombinant IL-33 (ACROBiosystems) or 100 ng/ml bovine serum albumin (Gibco, USA) intervention. After 2 h of NF-κB inhibitor JSH-23 (purity ≥ 98%, 20 μM; Cayman), wortmannin (WM; 25 μg/ml; Sigma), and U0126 (10 μM; Promega) intervention, UVB and IL-33 interventions were carried out.
8
Inhibitors for Cell Signaling Studies
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17β-Estradiol (E2) and PI3K inhibitor Wortmannin (WM) were purchased from Sigma-Aldrich (Milan, Italy). G-1 (1-[4-(− 6-bromobenzol [1,3]diodo-5-yl)-3a,4,5,9b-tetrahidro3H5cyclopenta[c]quinolin-8yl]-ethanone) and G-15 (3aS, 4R, 9bR)-4-(6-bromo-1, 3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta [c] quinolone were obtained from Tocris Bioscience (Space, Milan, Italy). Src kinase inhibitor PP2 was bought from Selleckchem (DBA, Milan, Italy). MEK inhibitor PD98059 (PD) was purchased from Calbiochem (DBA, Milan, Italy). STAT3 transcription factor signaling inhibitor STA21 and Focal Adhesion Kinase selective inhibitor VS-4718 were bought from Santa Cruz Biotechnology (DBA, Milan, Italy). All the aforementioned compounds were dissolved in dimethyl-sulfoxide (DMSO).
9
Tomato Peptide Signaling Pathway
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The tomato (Solanum lycopersicum) cultivar condine red and N. benthamiana were used in this study. Germinated seeds were grown in 100 cm 3 plastic pots (one plant per pot) containing peat and vermiculite (7:3, v/v), receiving Hoagland nutrient solution. The growth conditions were maintained at 23/21 °C (day/night) and a photoperiod of 12 h with 400 µmol m -2 s -1 photosynthetic photon flux density.
Five-wk-old tomato plants were sprayed with 10 μM PSK (Iris Biotech) or dH 2 O as control 12 h before pathogen inoculation, and N. benthamiana leaves were infiltrated with 1 μM PSK (Iris Biotech) or dH 2 O as control 1 h before sample collection. For the protein abundance assay, the uppermost one or two fully expanded leaves from 5-wk-old OE-PSKR1 tomato plants were inoculated with 1 μM PSK, 200 μM CHX (MedChemExpress), 50 μM MG132 (MedChemExpress), 33 μM wortmannin (Wm) (Sigma-Aldrich), 0.1 μM CMA (MedChemExpress), and the corresponding control dH 2 O or dimethyl sulfoxide (DMSO).
Five-wk-old tomato plants were sprayed with 10 μM PSK (Iris Biotech) or dH 2 O as control 12 h before pathogen inoculation, and N. benthamiana leaves were infiltrated with 1 μM PSK (Iris Biotech) or dH 2 O as control 1 h before sample collection. For the protein abundance assay, the uppermost one or two fully expanded leaves from 5-wk-old OE-PSKR1 tomato plants were inoculated with 1 μM PSK, 200 μM CHX (MedChemExpress), 50 μM MG132 (MedChemExpress), 33 μM wortmannin (Wm) (Sigma-Aldrich), 0.1 μM CMA (MedChemExpress), and the corresponding control dH 2 O or dimethyl sulfoxide (DMSO).
10
Pharmacological Modulation of COX-2 and EP4 Signaling
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NS398 (COX-2 inhibitor) was purchased from Cayman Chemical (Michigan, MI). ONO-AE3-208 (selective EP4 antagonist) was a gift of ONO Pharmaceuticals, Osaka, Japan. PGE2 (EP1-4 ligand) and PGE1OH (EP4 agonist) were purchased from Cayman Chemical. Wortmannin (WM), an irreversible PI3K inhibitor, and LY-204002 (LY), a reversible PI3K inhibitor all purchased from Sigma-Aldrich, ERK1/2 inhibitor U0126 from Cell Signalling, MA. NF-κB inhibitor, Bay 11-7082 (Sigma-Aldrich, ON, Cat # B5556) is a kind gift of Dr. Xiufen Zheng (Department of Pathology and Laboratory Medicine, University of Western Ontario). For all treatments in vitro or in vivo, respective vehicles served as the control.
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