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11 protocols using rottlerin

1

Formulation and Characterization of Nanoparticles

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FAM powder, polyvinylpyrrolidone (PVP), gum arabic, isoflurane, D-mannitol, methyl p-hydroxybenzoate, and cytochalasin D were purchased from Wako Pure Chemical Industries Ltd. (Osaka, Japan). The Bio-Rad Protein Assay Kit and 2-hydroxypropyl-β-cyclodextrin (HPβCD) were obtained from Bio-Rad (CA, United States) and Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan), respectively. Nystatin was purchased from Sigma–Aldrich (St. Louis, MO, United States). Pentobarbital and 1-heptanesulfonic acid sodium salt were obtained from Tokyo Chemical Industry Co. Ltd. (Tokyo, Japan). Methylcellulose (MC) was purchased from Shin-Etsu Chemical Co. Ltd. (Tokyo, Japan). Rottlerin and dynasore were obtained from Nacalai Tesque (Kyoto, Japan). All chemicals used were of the highest commercially available purity.
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2

Nanomaterial-Based Drug Delivery Protocol

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Indomethacin powder (particle size, 5.01±0.59 µm), cytochalasin D, isoflurane, mannitol (D-mannitol) and propyl p-hydroxybenzoate were purchased from Wako Pure Chemical Industries, Ltd (Osaka, Japan). 2-Hydroxypropyl-β-cyclodextrin (HPβCD) was obtained from Nihon Shokuhin Kako Co., Ltd (Tokyo, Japan), and nystatin and rat tail collagen type 1, was provided by Sigma-Aldrich Japan (Tokyo, Japan). Methylcellulose (MC, type SM-4) with an average viscosity of ~4 Pa·s at 20°C was obtained from Shin-Etsu Chemical Co., Ltd (Tokyo, Japan). Dynasore and rottlerin were purchased from Nacalai Tesque (Kyoto, Japan), and benzalkonium chloride (BAC) was provided by Kanto Chemical Co., Inc. (Tokyo, Japan). Dulbecco’s Modified Eagle’s Medium/Ham’s F12, heat-inactivated fetal bovine serum, streptomycin and penicillin were obtained from Gibco (Thermo Fisher Scientific, Waltham, MA, USA). TetraColor One was purchased from Seikagaku Co. (Tokyo, Japan). All other chemicals used were of the highest purity commercially available.
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3

Topical Delivery of Terbinafine Hydrochloride

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Pentobarbital was purchased from Sumitomo Dainippon Pharma Co., Ltd. (Toyo, Japan). TUL, cytochalasin D, methyl p-hydroxybenzoate, polyethylene glycol (PEG) 4000, cetyl alcohol, l-menthol, mineral oil, white wax, sodium tetraborate, propylene glycol, beeswax, and sodium dodecyl sulfate were provided by Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Commercially available 0.5 mg TUL tape (CA-TUL, Hokunalin® Tapes 0.5 mg) was obtained from Mylan EPD G.K (Tokyo, Japan). Rottlerin and dynasore were purchased from Nacalai Tesque (Kyoto, Japan), and methylcellulose (MC, SM-4) was obtained from Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan). 2-hydroxypropyl-β-cyclodextrin (HPβCD) was supplied by Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan), and PEG 400 was provided by Maruishi Pharmaceutical Co., Ltd. (Osaka, Japan). Nystatin and 450 nm pore size MFTM membrane filters were purchased from Sigma-Aldrich Japan (Tokyo, Japan) and Merck Millipore (Tokyo, Japan), respectively. All other chemicals used were of the highest purity commercially available.
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4

Comprehensive Reagents for Cell Culture

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Dulbecco’s Modified Eagle’s Medium (high-glucose, DMEM), non-essential amino acids solution, rottlerin, and dynasore were purchased from Nacalai Tesque (Kyoto, Japan), and heat-inactivated fetal bovine serum, penicillin, and streptomycin were obtained from Grand Island Biological Company (GIBCO, Tokyo, Japan). l-glutamine and nystatin were purchased from Sigma-Aldrich Japan (Tokyo, Japan). Indomethacin powder (particle size, 16.3 ± 12.9 μm), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, propyl p-hydroxybenzoate, isoflurane, and cytochalasin D were provided by Wako Pure Chemical Industries, Ltd. (Osaka, Japan). HPβCD and MC (type SM-4) were supplied by Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan) and Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan), respectively. Pentobarbital was obtained from Tokyo Chemical Industry Co., Ltd (Tokyo, Japan), and methanol and acetonitrile were provided by Kanto Chemical Co., Inc. (Tokyo, Japan). The Bio-Rad Protein Assay Kit was purchased from Bio-Rad Laboratories, Inc. (Bio-Rad, CA, U.S.A.). All other chemicals were of the highest purity which was commercially available.
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5

Microcellular Delivery of Indometacin

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All reagents used were the highest purity commercially available. Briefly, the HPβCD was provided by Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan), and MC was obtained from Shin-Etsu Cheical Co., Ltd. (Tokyo, Japan). IND powder, l-menthol, isoflurane, cytochalasin D, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, and propyl p-hydroxybenzoatewere were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Rottlerin (micropinocytosis inhibitor) and dynasore (CME inhibitor) were obtained from Nacalai Tesque (Kyoto, Japan). Carbopol (carboxypolymethylene, Carbopol® 934) and nystatin were purchased from Serva (Heidelberg, Germany) and Sigma–Aldrich Japan (Tokyo), respectively.
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6

Comprehensive Reagent Sourcing for Cell-based Assays

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Lan, rottlerin, dynasore, and Cell Count Reagent SF were purchased from Nacalai Tesque, Inc. (Kyoto, Japan), and a lipid peroxidation (LPO) assay kit (BIOXYTECH® LPO-586TM) was provided by OXIS International, Inc. (Portland, Oregon, USA). Nystatin was obtained from Sigma-Aldrich Japan (Tokyo, Japan). Heat-inactivated fetal bovine serum, penicillin, streptomycin, and Dulbecco’s modified Eagle’s medium/Ham’s F12 (DMEM/F12) were purchased from GIBCO (Tokyo, Japan). Benzalkonium chloride (BAC) and a Bio-Rad protein assay kit were obtained from Kanto Chemical Co., Inc. (Tokyo, Japan) and Bio-Rad Laboratories (Hercules, CA, USA), respectively. Ca test kits, mannitol, cytochalasin D, and isoflurane were obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan), and 0.1% pivalephrine and 0.4% benoxil were purchased from Santen Pharmaceutical Co., Ltd. (Osaka, Japan). MC (type SM-4) and HPCD was supplied by Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan) and Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan), respectively. SUPER FIX was provided by Kurabo Industries (Osaka, Japan). Calpain Activity Fluorometric Assay Kits were purchased from BioVision Inc. (San Francisco, CA, USA).
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7

Comprehensive Compound Acquisition for Research

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FL powder, LV powder, cytochalasin D, isoflurane, mannitol (D-mannitol) and butyl p-hydroxybenzoate were purchased from Wako Pure Chemical Industries, Ltd (Osaka, Japan). 2-hydroxypropyl-β-cyclodextrin (HPβCD) was obtained from Nihon Shokuhin Kako Co., Ltd (Tokyo, Japan), and nystatin was provided by Sigma-Aldrich Japan (Tokyo, Japan). Methylcellulose (MC) was obtained from Shin-Etsu Chemical Co., Ltd (Tokyo, Japan). BF, dimethyl sulfoxide (DMSO), dynasore, rottlerin and Cell Count Reagent SF were purchased from Nacalai Tesque (Kyoto, Japan), and BAC was provided by Kanto Chemical Co., Inc. (Tokyo, Japan). Fetal bovine serum, Dulbecco’s modified Eagle’s medium/Ham’s F12 (DMEM/F12), streptomycin and penicillin were provided by GIBCO (Tokyo, Japan). CA-FL, CA-LV and CA-oxybuprocaine hydrochloride eye drops (Benoxil® ophthalmic solution 0.4%) were obtained from Santen Pharmaceutical Co., Ltd. (Osaka, Japan), and CA-BF was purchased from Nitto Medic Co., Ltd. (Toyama, Japan). All other chemicals used were of the highest purity commercially available.
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8

Preparation and Characterization of Irbesartan Powder

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Irbesartan powder (IRB-P), propyl p-hydroxybenzoate, and cytochalasin D were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Rottlerin and dynasore were provided from Nacalai Tesque (Kyoto, Japan), and nystatin was obtained from Sigma-Aldrich Japan (Tokyo, Japan). A Bio-Rad Protein Assay Kit was purchased from BIO-RAD (Hercules, CA, USA). Pentobarbital and methylcellulose (MC) were obtained from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan), and Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan), respectively. 2-Hydroxypropyl-β-cyclodextrin (HPβCD) was provided by Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan). All other chemicals were of the highest purity commercially available.
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9

Raloxifene Powder Formulation Development

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Conventional raloxifene powder (particle size, 4.91±0.31 µm, mean ± SD), l-menthol (menthol), isoflurane, methyl p-hydroxybenzoate, and cytochalasin D were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Carbopol® 934 (carbopol and carboxypolymethylene) and nystatin were provided from Serva (Heidelberg, Germany), and Sigma-Aldrich Japan (Tokyo, Japan) respectively. Methylcellulose (MC), obtained from Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan), had an average viscosity of ~4 Pa·s at 20°C. Dynasore and rottlerin were purchased from Nacalai Tesque (Kyoto, Japan). All other chemicals were used with the highest purity commercially available.
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10

Formulation and Characterization of Nystatin-Loaded Carbopol Hydrogel

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Nystatin and carboxypolymethylene (carbopol, Carbopol® 934) were purchased from Sigma-Aldrich Japan (Tokyo, Japan) and Serva (Heidelberg, Germany), respectively. Cytochalasin D, propyl p-hydroxybenzoate, isoflurane and commercially available ketoprofen powder (particle size, 7.7 ± 0.28 μm, means ± S.E.) were provided by Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Rottlerin and dynasore were purchased from Nacalai Tesque (Kyoto, Japan) and methylcellulose (MC, SM-4) was obtained from Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan). All other chemicals used were of the highest purity commercially available.
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