Rabbit polyclonal anti-myc antibody, mouse monoclonal anti-actinin-1 antibody, and DMSO were from Sigma–Aldrich (St Louis, MO, USA). Mouse monoclonal anti-Stx6 antibody was from BD Transduction Laboratories (San Jose, CA, USA). Rabbit polyclonal anti-Stx6 and anti-Stx16 antibodies were from Synaptic Systems (Goettingen, Germany). Mouse monoclonal anti-Tubulin antibody was from Abcam (Cambridge, MA, USA). Human holo-transferrin conjugated to A488 was from Invitrogen (Grand Island, NY, USA). Mouse anti-myc (c-Myc 9E10) and rabbit anti-furin (H-220) were from Santa Cruz Biotechnology (Dallas, TX, USA). Polyclonal anti-P-Akt(308) and P-Akt(473) were obtained from Cell Signaling Technology (Danvers, MA, USA). Cy3- and A488-conjugated donkey anti-rabbit and donkey anti-mouse secondary antibodies and horseradish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). Nocodazole was purchased from EMD Biosciences Inc. (Darmstadt, Germany) (10 mM stock in DMSO) and C2-ceramide was purchased from Enzo Life Sciences (Farmingdale, NY, USA) (50 mM stock in DMSO). Pre-designed siRNA for Stx6 (siStx6: 5′-CCGAGTCATCAGAAGAACTAA-3′) and non-related (siNR: 5′-AATAAGGCTATGAAGAGATA C-3′) were from Qiagen (Valencia, CA, USA). Human insulin was purchased from Eli Lilly (Indianapolis, IN, USA).
Dimethyl sulfoxide (dmso)
Manufactured by Enzo Life Sciences
Sourced in United States
DMSO (Dimethyl Sulfoxide) is a colorless, polar, aprotic solvent commonly used in various laboratory applications. It has a high boiling point and is miscible with water and many organic solvents. DMSO is often utilized as a solvent or cryoprotectant in cell and tissue culture, as well as for sample preparation and preservation in analytical techniques.
Automatically generated - may contain errors
Lab products found in correlation
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Diphenyleneiodonium, by Enzo Life Sciences (1 mentions)
Vivatome system, by Zeiss (1 mentions)
Chloroquine, by Enzo Life Sciences (1 mentions)
Rapamycin, by Enzo Life Sciences (1 mentions)
Earle s balanced salt solution ebss, by Merck Group (1 mentions)
Staurosporine, by Enzo Life Sciences (1 mentions)
Erlotinib, by Enzo Life Sciences (1 mentions)
Penicillin, by Euroclone (1 mentions)
Streptomycin, by Euroclone (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Euroclone (1 mentions)
Fibronectin tetrapeptide, by Santa Cruz Biotechnology (1 mentions)
Dasatinib s1021, by Selleck Chemicals (1 mentions)
Saracatinib s1006, by Selleck Chemicals (1 mentions)
Anti cd3 and anti cd28 antibody, by Thermo Fisher Scientific (1 mentions)
Human cd4 microbeads, by Miltenyi Biotec (1 mentions)
Anti cd3 cd28, by Thermo Fisher Scientific (1 mentions)
Luteolin, by Enzo Life Sciences (1 mentions)
9 protocols using dimethyl sulfoxide (dmso)
1
Characterizing Protein Trafficking Mechanisms
2
Zebrafish Hindbrain Infection Assay
3
Glucose Deprivation and Autophagy Modulation
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For glucose starvation, INS(832/13) and islet cells were grown for 24 h in complete RPMI1640 medium containing 2.8 mmol/l glucose (low glucose, LG). Controls/non-treated (NT) INS(832/13) cells were grown in complete RPMI1640 medium containing 11.1 mmol/l glucose. Cells were also incubated with 0.5 μmol/l rapamycin, dissolved in 0.04% DMSO (an autophagy inducer; Enzo, Plymouth Meeting, PA, USA [24 h incubation]), 10 μmol/l chloroquine (a lysosomal inhibitor; Enzo [24 h incubation]) or in amino-acid- and serum-free buffer (Earle’s Balanced Salt Solution [EBSS], Sigma Aldrich [4 h incubation]).
4
Erlotinib Cytotoxicity Experimental Protocol
5
Cell lines and culture conditions for DNA repair study
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HEK293T cells stably expressing Flag-tagged FUS and HeLa FUS-KO were described in Reber et al. (2016) (link). U2OS cells stably expressing HR and NHEJ repair reporters were a kind gift from J.M. Stark (Beckman Research Institute of City of Hope, Duarte, CA) and were described in Gunn and Stark (2012) (link). HeLa and SH-SY-5Y FUS-KO cells were generated by CRISPR-trap (Reber et al., 2018 (link)). All cell lines were tested for mycoplasma contamination and were found to be negative. Cells were cultured in DMEM (high glucose) supplemented with 10% FBS, 2 mM l -glutamine, 100 IU/ml penicillin, and 0.1 mg/ml streptomycin (all EuroClone). Cells were grown at 37°C and 5% CO2 in a humidified incubator.
ETO treatment, where not specified otherwise, was performed for 1 h with 10 µM diluted in DMSO (Enzo Lifesciences). Where specified, cells were treated with 2% 1,6-HD (diluted in growth medium; Sigma-Aldrich), 2% 2,5-HD (diluted in growth medium; Sigma Aldrich), or 50 or 100 mM Am. Ac. (diluted in water; Sigma Aldrich) for 30 min.
Plasmid DNA transfections were performed using Lipofectamine 2000 (Invitrogen), while siRNA transfections were done using Lipofectamine RNAiMAX (Invitrogen), according to the manufacturer’s instructions. The synthetic siRNAs used in this study were purchased from Riboxx Life Sciences.
ETO treatment, where not specified otherwise, was performed for 1 h with 10 µM diluted in DMSO (Enzo Lifesciences). Where specified, cells were treated with 2% 1,6-HD (diluted in growth medium; Sigma-Aldrich), 2% 2,5-HD (diluted in growth medium; Sigma Aldrich), or 50 or 100 mM Am. Ac. (diluted in water; Sigma Aldrich) for 30 min.
Plasmid DNA transfections were performed using Lipofectamine 2000 (Invitrogen), while siRNA transfections were done using Lipofectamine RNAiMAX (Invitrogen), according to the manufacturer’s instructions. The synthetic siRNAs used in this study were purchased from Riboxx Life Sciences.
6
Screening FDA-Approved Drug Library
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The library of FDA-approved drugs (Screen-Well FDA-Approved Drug Library, 640 chemical compounds dissolved at 10 mM in dimethylsulphoxide (DMSO)) was obtained from Enzo Life Sciences (Enzo Life Sciences Inc.,Plymouth Meeting, PA, USA).
The following compounds were purchased from Sigma Aldrich: HC (H0888), verteporfin (SML0534), PX (T7191), RU486 (Mifepristone; M8046), BM (B7005), PF573228 (PZ0117). RGD is fibronectin tetrapeptide (H-Arg-Gly-Asp-Ser-OH) from Santa Cruz (sc-202156).
Latrunculin A (sc-202691) was from Sigma, saracatinib (S1006) and dasatinib (S1021) from Selleck.
8XGTII-Luc and the retroviral construct coding for Flag-YAP-5SA were previously described16 (link). Lentiviral constructs coding for short hairpin NR3C1 (GR) was from Sigma (Clone ID:NM_000176.2-6190s21c1).
The following compounds were purchased from Sigma Aldrich: HC (H0888), verteporfin (SML0534), PX (T7191), RU486 (Mifepristone; M8046), BM (B7005), PF573228 (PZ0117). RGD is fibronectin tetrapeptide (H-Arg-Gly-Asp-Ser-OH) from Santa Cruz (sc-202156).
Latrunculin A (sc-202691) was from Sigma, saracatinib (S1006) and dasatinib (S1021) from Selleck.
8XGTII-Luc and the retroviral construct coding for Flag-YAP-5SA were previously described16 (link). Lentiviral constructs coding for short hairpin NR3C1 (GR) was from Sigma (Clone ID:NM_000176.2-6190s21c1).
7
Isolation and Activation of CD4+ T cells
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Peripheral blood samples were obtained from BD patients and healthy volunteers. Preparation of PBMCs and CD4+T cells was performed as described earlier [23 (link)]. Human CD4+T cells were isolated from PBMCs by human CD4 microbeads (purity > 90%; Miltenyi Biotec, Palo Alto, CA) according to the manufacturer's instructions. The PBMCs and purified CD4+T cells were cultured at 1 × 106 in 24-well plates in RPMI1640 medium supplemented with 10% FBS. A combination of anti-CD3 and anti-CD28 antibodies (2 ug/mL) (eBioscience, San Diego, Calif) was used to activate PBMCs for 3 days, and a combination of anti-CD3/CD28 (2 ug/mL) and IL-23 was used to activate CD4+T cells for 6 days. When used, 0.05% DMSO (control), FICZ (100 nmol/L) (Enzo Life Sciences, USA), and ITE (100 nmol/L, Tocris Bioscience, USA) were added at the beginning of the culture. The supernatants were collected for cytokine measurement by ELISA and the cells were harvested and used for FACS analysis or mRNA quantification.
8
Flavone Materials and Recombinant DGAS
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Nine flavone materials (apigenin, chrysin, 6,7-dihydroxyflavone, homoorientin, 7-hydroxyflavone, isorhoifolin, luteolin, luteolin-3′,7-diglucoside, and orientin; each prepared with 2 mg/mL DMSO) were purchased from Enzo Life Sciences (Farmingdale, NY, USA). The recombinant DGAS was prepared according to previous methods [25 ]. All other chemicals used in this study were of analytical reagent grade and were obtained from Sigma-Aldrich (Merck, Darmstadt, Germany).
9
Hydrogel Matrix Modulates hVSMC Response
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Serum-starved hVSMCs were plated on fibronectin-coated soft or stiff hydrogels with medium containing 10% FBS and were treated with 0.1, 0.5, or 2 μM YM155 (survivin inhibitor; Cat. No. 11490, Cayman Chemical), 10 μM PF573228 (FAK inhibitor; Cat. No. 14924, Cayman Chemical), 5 or 30 μM mitomycin C (proliferation inhibitor; Cat. No. BML-GR311-0002, Enzo Life Sciences), or DMSO (vehicle control).
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