0.2–0.3 × 106 iPSCs (OAS1-WT and OAS1-MUT) were subjected to terminal differentiation to macrophages in 24 well plates. Curcumin (FUJIFILM Wako Pure Chemical Corporation, Tokyo, Japan) was added to the culture at 25mM at 24 hours after initiation of differentiation and incubation was continued for another 24 hours. The cells were then cultured in the presence of IFN-α (1,000 IU/mL, R&D Systems, Minneapolis, USA) for 48 hours, at the end of which floating cells in the culture medium and viable adherent cells were counted under by microscopy. Cells were cultured in duplicate and the ratio of floating single cells (FSCs) and viable adherent cells were calculated for three independent experiments.
Curcumin
Manufactured by Fujifilm
Sourced in Japan
Curcumin is a laboratory equipment product manufactured by Fujifilm. It is a pure, highly concentrated curcuminoid compound extracted from the turmeric plant. Curcumin is used as a standard reference material in various analytical and research applications.
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Lab products found in correlation
Dimethyl sulfoxide (dmso), by Fujifilm (4 mentions)
Quercetin, by Fujifilm (2 mentions)
Ifn α, by R&D Systems (1 mentions)
Caffeine, by Fujifilm (1 mentions)
N acetylcysteine nac, by Merck Group (1 mentions)
Reproneuro, by ReproCELL (1 mentions)
Paraquat, by Fujifilm (1 mentions)
Horseradish peroxidase hrp conjugated anti mouse and anti rabbit secondary antibodies, by Cell Signaling Technology (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Epigallocatechin, by Merck Group (1 mentions)
Daidzein, by Fujifilm (1 mentions)
Apigenin, by Fujifilm (1 mentions)
Resveratrol, by Fujifilm (1 mentions)
Clodronate, by Merck Group (1 mentions)
Chrysin, by Fujifilm (1 mentions)
Flavanone, by Fujifilm (1 mentions)
Flavone, by Fujifilm (1 mentions)
Genistein, by Merck Group (1 mentions)
Ursolic acid, by Fujifilm (1 mentions)
Carnosol, by Fujifilm (1 mentions)
10 protocols using curcumin
1
Curcumin Modulates IFN-α Response in Macrophages
2
Neuronal Cell Culture and Toxicity Assays
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SH-SY5Y (ECACC, Salisbury, Wiltshire, United Kingdom), HCT-15, HCT116, HEK293, HEK293T and MCF-7 cells were cultured in DMEM/F-12 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum. To obtain normal human matured neuronal cells in a culture system, ReproNeuro, a neuron progenitor derived from human iPS cells, was purchased from ReproCELL (Yokohama, Japan) and maintained in ReproNeuro maturation medium for 14 days according to the manufacturer’s instructions. Carbonyl cyanide m-chlorophenyl hydrazone (CCCP), 6-hydroxydopamine hydrobromide (6-OHDA), cadmium chloride (CdCl2), rotenone, tert-Butylhydroquinone (tBHQ), sulforaphane and N-acetylcysteine (NAC) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Paraquat, caffeine and curcumin were purchased from Wako Chemicals (Osaka, Japan). The antibodies used were as follows: an antibody against PINK1 (rabbit monoclonal, 6946), an antibody against NRF2 (rabbit monoclonal, 12721), horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit secondary antibodies (Cell Signaling Technologies, Danvers, MA, USA), and an antibody against Tubulin (mouse monoclonal, T5168; Sigma-Aldrich).
3
Cell Proliferation Assays of Food Constituents
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We examined the inhibitory activities of food constituents in cell proliferation assays. Test compounds used in the assays were as follows: curcumin (Wako or Sigma-Aldrich), zerumbone (Wako), genistein (Sigma-Aldrich), chalcone (Wako), corosolic acid (Wako), carnosol (Wako), ursolic acid (Wako), epigallocatechin (Sigma-Aldrich), isoflavone (Wako), apigenin (Wako), chrysin (Wako), flavanone (Wako), resveratrol (Wako), flavone (Wako), quercetin (Wako), daidzein (Wako), and clodronate (Sigma-Aldrich). These compounds were firstly dissolved with dimethyl sulfoxide (DMSO; Wako) and then appropriately diluted with RPMI1640 + 10% FCS to apply the assays. The final concentration of DMSO was adjusted not to exceed 1% at most in cell cultures.
4
Synthesis and Evaluation of Curcumin Analogs
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GO‐Y078, GO‐Y030, GO‐Y022, and GO‐Y136 were synthesized as previously described21 , 22 , 23 (Figure 1 ). Wako special grade Curcumin was purchased from Wako Pure Chemical Industries, Ltd (Osaka, Japan). Curcumin and its analogs were dissolved in DMSO (Wako Pure Chemical Industries, Ltd) and diluted at final concentrations of 0.002%‐0.1%. Sunitinib, sorafenib, and Ki8751, which are selective VEGFR‐2 inhibitors, were purchased from Sigma‐Aldrich (St Louis, MO, USA), LC Laboratories (Woburn, MA, USA), and Wako Pure Chemical Industries, Ltd, respectively.
5
Degradation of Dyes by B. multivorans CCA53
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CCA53 was previously deposited in the HUT culture collection under strain number HUT-8135. Bordeaux S, bromothymol blue, congo red, cresol red, curcumin, ethyl red, fluorescein, methyl orange, methyl red, orange II, rhodamine B, thymol blue, and trypan blue were purchased from Fujifilm Wako Chemicals (Osaka, Japan) or Tokyo Chemical Industry (Tokyo, Japan). To confirm dyedegradation capacity, B. multivorans CCA53 was aerobically grown on LB plates (pH 7.0) containing 100 µM each dye for 3 days at 30°C.
6
Bisacurone Inhibits Lipid Accumulation
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Bisacurone was purchased from Nagara Science Co., Ltd. (Gifu, Japan). Curcumin, palmitic acid, oleic acid, and ImmunoStar LD was purchased from FUJIFILM Wako Pure Chemical Industries (Osaka, Japan). Their chemical structures are shown in Fig. 1 . Metformin hydrochloride was from Sigma-Aldrich (St. Louis, MO). Dulbecco’s modified Eagle’s medium (DMEM) was obtained from Nissui Pharmaceutical (Tokyo, Japan). Bovine serum albumin (BSA), Blocking One, and Blocking One-P solutions were from Nacalai Tesque, Inc. (Kyoto, Japan). For western blotting, antibodies for lamin B, PPARα, CPT1, fatty acid binding protein 4 (FABP4), C/EBPα, FAS, anti-mouse IgG, anti-goat IgG, and anti-rabbit IgG were purchased from Santa Cruz Biotechnology (Santa Cruz Biotechnology, Santa Cruz, CA); β-actin, p-AMPKα, AMPKα, p-LKB1, and LKB1 were from Cell Signaling Technology (Beverly, MA); ChREBP and SREBP-1 were from Abcam (Cambridge, MA). The polyvinylidene difluoride membrane was products of GE Healthcare Bio-Science Co., (Piscataway, NJ). All other reagents used were of the highest grade available from commercial sources.
7
Curcumin Compound Preparation for Research
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Curcumin (Wako, Japan) stock solutions were prepared in sterile dimethyl sulfoxide (DMSO). The Curcumin analogue GO-Y030 was provided by Akita University, Japan. Its stock solution was prepared in sterile DMSO and stored at 4°C. Nile red (Wako, Japan) stock solution was prepared in ethanol and stored at 4°C. Doxycycline (Wako, Japan) was prepared in distilled water and stored at 4°C. Curcumin compounds and all dye solutions were kept in the dark to prevent light exposure.
8
Investigating NF-κB Inhibitors in Oxaliplatin and Curcumin Treatment
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Oxaliplatin and curcumin were purchased from FUJIFILM Wako Pure Chemical. Three known NF‐κB inhibitors (Bay11‐7082; IκBα kinase inhibitor, TPCA1; IκB kinase inhibitor, MG132; proteasome inhibitor) were purchased from Abcam for Bay11‐7082 and TPCA1, or from Cayman Chemical for MG132, respectively. CMG was synthesized using a previous method (KNC Laboratories).21
9
Lipid Nanoparticle Formulation and Characterization
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Coconut oil, PEG600, Tween 80 (T80), cholesterol and 2-deoxy-D-glucose were purchased from Sigma-Aldrich (St. Louis, MO). Egg phosphatidylcholine (EPC) was obtained from NOF (Tokyo, Japan). Sodium Azide, Curcumin, Dulbecco's Modified Eagle's Medium (DMEM), Dimethyl sulfoxide (DMSO) and Diisopropyl ether were obtained from Wako Chemicals (Osaka, Japan). The 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine (DiD) fluoresce Probe was supplied by Thermo Fisher Scientific (Waltham, MA), and both NBD-DOPE and Rhodamine-DOPE were purchased from Avanti Polar Lipids (Alabaster, Al). A Cell Counting Kit-8 was obtained from Dojindo (Kumamoto, Japan) and RC Dialysis membrane (MWCO 12-14 kD) from Spectrum Laboratories (Rancho Dominguez, CA). Monolein and Triolein were purchased from Kanto chemicals and TCI, Respectively (Tokyo, Japan).
10
Comparative Analysis of Bioactive Compounds
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The following 10 chemicals were tested in our study, with relevant information summarized in Table 1: carminic acid, esculetin, 4-Methyl esculetin (purchased from Tokyo Kasei, Tokyo Chemical Industry Co. Ltd., Tokyo, Japan); coumarin, quercetin, curcumin, naringenin, chlorogenic acid acetone and dimethyl sulfoxide (DMSO) (purchased from Wako, Wako Pure Chemical Industries, Ltd., Osaka, Japan); isoscopoletin (purchased from Funakoshi, Funakoshi Co. Ltd., Tokyo, Japan); and shikonin (purchased from Kishida, Kishida Chemical Co. Ltd., Osaka, Japan). Olive oil used for testing was purchased from Yoshida Pharmaceutical Co. Ltd., Japan.
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