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2 protocols using ascomycin

1

Silymarin and Irinotecan Formulation Study

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Silymarin (80%) was purchased from Sanjaing (Jiaxing, China). Irinotecan hydrochloride and SN-38 were purchased from Scino Pharm (Tainan, Taiwan). SN38G was purchased from Cayman Chemical (Ann Arbor, MI, USA). Rapamycin was purchased from Chunghwa Chemical Synthesis and Biotech (New Taipei City, Taiwan). Capryol-90 was purchased from Gattefosse (Lyon, France). Tween 80 and camptothecin were purchased from Merck KGaA (Darmstadt, Germany). Cremophor EL was procured from Wei Ming Pharmaceutical (Taipei, Taiwan). Ascomycin was purchased from MedChemExpress (South Brunswick, NJ, USA). Soybean lecithin (Lipoid S-100) was purchased from Lipoid GmbH (Ludwigshafen, Germany). Dulbecco’s modified Eagle’s medium, fetal bovine serum, and horse serum were purchased from Corning (New York, NY, USA). Reagents used for high-performance liquid chromatography (HPLC) or ultra-performance liquid chromatography with tandem mass spectrometry (UPLC/MS/MS) were of HPLC or MS grade, and other reagents were of analytical grade.
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2

Chemical Treatments in N. benthamiana

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N. benthamiana leaves at 48 h after infiltration were treated with 1 μM rapamycin (Sigma-Aldrich) and/or 50 μM oryzalin (Sigma-Aldrich) or 250 nM MitoTracker™ Red CMXRos (Thermo Fisher) and/or 100 μM ascomycin (MedChemExpress) by syringe-infiltration. Stock solutions (1 mM rapamycin, 20 mM oryzalin, 1 mM MitoTracker and 10 mM ascomycin) were prepared by diluting the corresponding amount of chemical in DMSO (dimethyl sulfoxide). Prior to infiltration, the chemicals were diluted to their final concentrations in MiliQ water.
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