The backs of the mice were shaved with electric clippers 1 day before PUVA treatment. Groups of mice were painted on their backs either with 100 μl of vehicle (100% ethanol) or 8-MOP (Sigma-Aldrich, Saint Louis, Missouri) in ethanol (at a concentration of 1 mg/ml), or were left untreated. The mice were then kept for 30 min in individual compartments of standard cages, separated with Plexiglass dividers to allow penetration of 8-MOP. UVA irradiation was provided by bank of six Sylvania F15T8/BLB lamps (emission range, 320 to 400 nm; peak at 365 nm; Danvers, MA). During UVA irradiation, the mice were housed five per cage, individually separated, on a shelf 20 cm below the fluorescent light bulbs under a wire cage top. The mice were UVA-irradiated at a mean UVA irradiance of 4.5 mW/cm2, as measured by an IL 700 spectroradiometer with a SEE 033 UVA detector (International Light Inc., Newburyport, MA) (3 (link)). UVB irradiation was produced by an Oriel solar simulator, as previously described (28 (link)).
8 mop
Manufactured by Merck Group
Sourced in United States
8-MOP is a laboratory reagent used in photochemical research and analysis. It functions as a photosensitizing agent, enhancing the sensitivity of samples to light exposure. The detailed technical specifications and intended applications of this product are not included in this factual description.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
5 mop, by Merck Group (2 mentions)
β nicotinamide adenine dinucleotide phosphate reduced form nadph, by Merck Group (2 mentions)
Caffeine, by Merck Group (2 mentions)
Acetonitrile, by Thermo Fisher Scientific (2 mentions)
Methanol, by Thermo Fisher Scientific (2 mentions)
Nitrogen gas, by Linde (1 mentions)
Fusidic acid, by Chem-Impex (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Rifampin, by MedChemExpress (1 mentions)
Chloramphenicol, by Merck Group (1 mentions)
Clindamycin, by Merck Group (1 mentions)
Niraparib, by Selleck Chemicals (1 mentions)
Talazoparib, by Selleck Chemicals (1 mentions)
Anti histone h3 antibody, by Abcam (1 mentions)
Anti β actin antibody, by Merck Group (1 mentions)
Anti parp2 antibody, by Active Motif (1 mentions)
Anti par antibody, by R&D Systems (1 mentions)
Olaparib, by Selleck Chemicals (1 mentions)
Anti α tubulin antibody, by Merck Group (1 mentions)
12 protocols using 8 mop
1
Photochemotherapy Protocol for Mice
2
Radiolabeled Caffeine Metabolism Analysis
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Caffeine (≥99.0 %), 8-MOP (≥98.0 %), 5-MOP (99.0 %), ISOP (≥95.0 %) and β-nicotinamide adenine dinucleotide phosphate reduced form (NADPH) (≥97.0 %) were purchased from Sigma-Aldrich (St. Louis, MO) and ChromaDex (Irvine, CA). Trichloroacetic acid (≥99.0), dipotassium phosphate (≥60.0), and monopotassium phosphate (≥60.0) were provided by Anachemia (Rouses Point, NY). Nitrogen gas (≥99.9 %) was procured from Praxair (Danbury, CT). Dimethyl sulfoxide (DMSO) (spectral grade) was provided by Caledon (Georgetown, ON). Radiolabeled Caffeine [3-methyl-14C] with specific activity of 50−60 mCi/mmol was purchased from American Radiolabeled Chemicals (St. Louis, MO). The scintillation cocktail fluids were obtained from PerkinElmer (Waltham, MA) and Amersham Biosciences (Piscataway, NJ). Ultrapure water was produced using a Millipore system (Billerica, MA) with a minimum resistivity of 16.0 MΩ cm at 25 °C.
3
Characterization of Psoralen-DNA Adducts
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Chemical reagents including HMT and 8-MOP were purchased from Sigma-Aldrich (France). All regular DNA oligonucleotides were purchased from Eurogentec (Seraing, Belgium). Sequences of all the oligonucleotides used in this work are described in Supplementary Table S2 . The 17-mer oligonucleotide d(CCACCAACSpCTACCACC) containing single spiroiminodihydantoin (Sp) lesion at position 9 was a gift from Dr Nicolas Geacintov (New York University, NY, USA) and was synthesized as described41 (link). Complementary oligonucleotides were hybridized to obtain duplexes referred to as D21∙C21, D21∙C47 and C47∙D101. Crosslinked D21-C47 and C47-D101 were hybridized with complementary D47 and C101 oligonucléotides to obtain three-stranded DNA structures XL47∙47-21 and XL101∙101-47. Crosslinked XL21-21 and XL47-21 oligonucleotide duplexes were used to generate MAs and ICLs. To obtain oligonucleotide containing single MAp residue, the denaturing gel purified crosslinked DNA duplexes were treated with hot alkali as described29 (link). Chemical structures of psoralen-DNA adducts used in this work are shown in Supplementary Table S1 .
4
Phototoxicity Assessment of Drugs
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The phototoxic drugs 8-Methoxy psoralen (8-MOP), Ibuprofen (IBF), Benzoyl peroxide (BPO), Ketoprofen (KPF), and Piroxicam (PXC) were used as positive substances and the non-phototoxic drug Sulisobenzone (SBZ) was used as a negative substance. A total of 6 substances were tested, and Acetone: DMSO (10:1) was used as the vehicle control (V.C). On the day of the test, test substances were prepared and the concentration of the substance was chosen as the highest concentration of soluble in vehicle control. 8-MOP, IBF, BPO, KPF, PXC, and SBZ were purchased from Sigma-Aldrich (St. Louis, MO, USA). All other reagents and materials were purchased from commercial sources.
5
Pathogen Reduction Reagent Preparation
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Amotosalen HCl 3 mM solution (lots CE19F14L71 and CE20J12L71) was obtained from the Cerus INTERCEPT Blood System for Platelets Pathogen Reduction System Dual Storage Processing Set and stored in light-protected aliquots at 4°C. AMT and 8-MOP, both from Sigma-Aldrich, were dissolved in ~ 4% dimethyl sulfoxide (DMSO) and 100% DMSO, respectively, and stored as aliquots at −80°C prior to use. Other antimicrobials used were apramycin (Alfa Aeser), clindamycin (Sigma-Aldrich), chloramphenicol (Sigma-Aldrich), fusidic acid (Chem-Impex International), gentamicin (Alfa Aeser), minocycline (Chem-Impex International), rifampin, and PAβN (MedChem Express).
6
PARP Inhibitor Evaluation Protocol
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The PARP inhibitors olaparib (Selleckchem, S1060), niraparib (Selleckchem, S2741), and talazoparib (Selleckchem, S7048) were dissolved in dimethyl sulfoxide (DMSO) and used at 1 μM final concentration. Anti-PARP1 antibody (CST, 9542) was used at 1:5000. Anti-PARP2 antibody (Active Motif, 39044) was used at 1:2000. Anti-XRCC1 antibody (Novus Biologicals, 87154) was used at 1:5000. Anti-PAR antibody (R&D, 4335-MC-100) was used at 1:1000. Anti-Histone H3 antibody (Abcam, ab1791) was used at 1:5000. Anti-α-Tubulin antibody (Sigma, CP06) was used at 1:5000. Anti-β-Actin antibody (Sigma, A5441) was used at 1:10 000. 8-MOP (Methoxsalen) (Sigma, M3501) was added 10 min before micro-irradiation at a final concentration of 100 μM when used.
7
Optimization of Liposomal Formulations
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8-MOP and Pluronic F68 (Poloxamer 188) were purchased from Sigma Aldrich (Milan, Italy). Compritol® 888 ATO and Transcutol® P (TRC) were kindly supplied by Gattefossè (Lyon, France). Cell culture materials were purchased from Invitrogen (Milan, Italy). Cholesterol, standards of fatty acids, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (PC 16:0/16:0), 1,2-dioleoyl-sn-glycero-3-phosphocholine (PC 18:1/18:1), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (PC 16:0/18:1), 1-oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine (PC 18:1/16:0), 2-linoleoyl-1-palmitoyl-sn-glycero-3-phosphocholine (PC 16:0/18:2), 2-arachidonoyl-1-palmitoyl-sn-glycero-3-phosphocholine (PC 16:0/20:4), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (PC 18:2/18:2), and 1,2-dieicosapentaenoyl-sn-glycero-3-phosphocholine (PC 20:5/20:5) were purchased from Sigma–Aldrich (Milan, Italy). All the other chemicals used in this study were of analytical grade.
8
Radiochemical Assay Reagents and Solvents
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Benzotriazole (99.0%), caffeine (≥99.0%), ethyl acetate (≥99.7%), 8-MOP (≥98.0%), 5-MOP (99.0%), and β-nicotinamide adenine dinucleotide phosphate reduced form (NADPH) (≥97.0%), were obtained from Sigma-Aldrich (St. Louis, MO). Methanol (≥99.9%) and acetonitrile (≥99.9%) were obtained from Thermo Fisher Scientific (Hampton, NH) and Sigma-Aldrich. Acetic acid (≥99.7%), trichloroacetic acid (TCA) (≥99.0), dipotassium phosphate (K2HPO4) (≥60.0), and monopotassium phosphate (KH2PO4) (≥60.0) were obtained from Anachemia (Rouses Point, NYC). ISOP was obtained from ChromaDex (Irvine, CA) (98.3%) and Sigma-Aldrich (≥95.0%). Spectral grade dimethyl sulfoxide (DMSO) was obtained from Caledon (Georgetown, ON). High-purity nitrogen gas (N2), oxygen gas (O2), and carbon monoxide (CO) gas was obtained from Praxair (Danbury, CT).
[3′-N-14C-methyl]-caffeine (specific activity 50–60 mCi/mmol) was obtained from American Radiolabeled Chemicals (St. Louis, MO). The methyl-14C labeled 8-MOP, specific activity of 40–60 mCi/mmol, was purchased from Vitrax Radiochemicals (Placentia, CA). Scintillation cocktail fluids were obtained from Perkin Elmer Life Sciences (Waltham, MA) and Amersham Biosciences (Piscataway, NJ). Ultrapure water was produced using a Millipore system (Billerica, MA) with a minimum resistivity of 16.0 MΩ cm at 25°C.
[3′-N-14C-methyl]-caffeine (specific activity 50–60 mCi/mmol) was obtained from American Radiolabeled Chemicals (St. Louis, MO). The methyl-14C labeled 8-MOP, specific activity of 40–60 mCi/mmol, was purchased from Vitrax Radiochemicals (Placentia, CA). Scintillation cocktail fluids were obtained from Perkin Elmer Life Sciences (Waltham, MA) and Amersham Biosciences (Piscataway, NJ). Ultrapure water was produced using a Millipore system (Billerica, MA) with a minimum resistivity of 16.0 MΩ cm at 25°C.
9
Adoptive Transfer of PUVA-Treated Spleen Cells
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Spleens of the donor mice (arthritic or healthy mice) were harvested and mechanically dissociated. Erythrocytes were removed with a red blood cell lysis solution (eBiosciences). Spleen cells (10·106 cells/ml) were treated by PUVA during 15 min at 37 °C with 8-MOP (200 ng/ml, Sigma-Aldrich) and irradiated with UV-A (365 nm, 2 J/cm2). Cells were washed twice with PBS. Irradiated cells (PUVA cells) were reinjected intravenously (10·106 cells in 200 µl of PBS) three times every other day. Treatment was initiated when the mean arthritic score reached 7. Mice receiving arthritic mice-derived PUVA-treated-spleen cells were called «Arthritic PUVA» (A-PUVA) mice while mice receiving healthy mice-derived PUVA treated-spleen cells were called «Healthy PUVA» (H-PUVA) mice.
10
Cell Lines and Reagents for Cancer Research
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General chemicals and reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA) unless stated otherwise. Gemcitabine and 8-MOP were purchased from Sigma-Aldrich. Inhibitors including Z-VAD-FMK, 3-MA, and necrostatin-1 were purchased from Selleckchem (Houston, TX, USA).
All human cancer cell lines were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea). Two cell lines, if available, were used for each type of cancer. The human cancer cell lines used in this study were as follows: A549 and H1650 (lung cancer); SNU449 and SK-Hep1 (hepatocellular carcinoma); MDA-MB453 and T47D (breast cancer); SKOV3 (ovarian cancer); PC3 and DU145 (prostate cancer); HCT15 and DLD1 (colorectal cancer); AGS and SNU1 (gastric cancer harboring wild-type p53); and SK-MEL-2 (skin cancer). Cells were cultivated in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% fetal bovine serum (FBS) (Thermo-Fisher Scientific, Waltham, MA, USA) and 1% antibiotic-antimycotic solution at 37°C in 5% CO2 atmosphere, unless stated otherwise.
All human cancer cell lines were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea). Two cell lines, if available, were used for each type of cancer. The human cancer cell lines used in this study were as follows: A549 and H1650 (lung cancer); SNU449 and SK-Hep1 (hepatocellular carcinoma); MDA-MB453 and T47D (breast cancer); SKOV3 (ovarian cancer); PC3 and DU145 (prostate cancer); HCT15 and DLD1 (colorectal cancer); AGS and SNU1 (gastric cancer harboring wild-type p53); and SK-MEL-2 (skin cancer). Cells were cultivated in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% fetal bovine serum (FBS) (Thermo-Fisher Scientific, Waltham, MA, USA) and 1% antibiotic-antimycotic solution at 37°C in 5% CO2 atmosphere, unless stated otherwise.
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