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5 protocols using salvianolic acid a

1

Antioxidant Compounds Cytotoxicity Evaluation

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Rosmarinic acid, salvianolic acid A, quercetin, and ursolic acid were purchased from Sigma-Aldrich/Merck (Algés, Portugal); luteolin-7-glucoside was obtained from Extrasynthese® (Genay, France); and eriodictyol and oleanolic acid were obtained from Santa Cruz Biotechnology Inc. (Frilabo, Porto, Portugal). Dulbecco’s modified Eagle medium (DMEM), versene, trypsin-EDTA, penicillin, streptomycin, fetal bovine serum (FBS), L-glutamine, and sodium pyruvate were obtained from Gibco (Alfagene, Lisboa, Portugal). Alamar Blue® was purchased from Invitrogen, Life-Technologies (Alfagene, Lisboa, Portugal). Dichlorodihydrofluorescein diacetate (DCFDA), 5,5″,6,6″-tetrachloro-1,1″,3,3″-tetraethylbenzimidazolylcarbocyanineiodide (JC-1), and Annexin-V-FITC were purchased from Thermo Fisher Scientific (Alfagene, Lisboa, Portugal). RNAse was obtained from NZYtech (Lisboa, Portugal). Flow cytometry solutions and consumables were obtained from BD—Becton Dickinson (Enzifarma, Lisboa, Portugal). Other salts and reagents not mentioned above were obtained from Sigma-Aldrich/Merck (Algés, Portugal).
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2

Identification of Antiviral Compounds from Small Molecule Libraries

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The Tocriscreen compound library (Tocris Bioscience), Pharmakon1600 drug library (Microsource Discovery Systems), and Prestwick chemical library (Prestwick chemical) were used for screening. The Pharmakon1600 drug library was generously provided by Dr. Koh Takeuchi (Advanced Industrial Science and Technology, Ibaraki, Japan). The Prestwick chemical library was kindly gifted by Dr. Masayuki Shimojima and Dr. Masayuki Saijo (National Institute of Infectious Diseases, Tokyo, Japan). Rosmarinic acid, quercetin, merbromin, hemin, calcomine orange 2RS, and salvianolic acid A were purchased from Sigma-Aldrich (536954, Q4951, M7011, 51280-1G, C9519, and SML0045, respectively). Erythrosine B, and lamivudine were purchased from Tokyo Chemical Industry (T0557 and L0217, respectively). Verteporfin was purchased from Cayman Chemical (17334). Rosmarinic acid derivatives were synthesized as previously described [28 (link)].
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3

NMNAT1 Regulation of Mitochondrial Function

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Salvianolic acid A (SML0045) was purchased from Sigma. High glucose DMEM (10569010), Lipofectamine 3000 Transfection Reagent (1L3000150), fetal bovine serum (FBS) (10099), G418 (10131027), and puromycin (A1113803) were purchased from Thermo Fisher. Total RNA Extraction kit (R1200), MTT reagent (M8180), and TaqMan One Step RT-qPCR Kit (T2210) were purchased from Solarbio. Quick Ligase (M2200S), HindIII (R3104S), and EcoRI (R3101S) were purchased from NEB. Esp3I (ER0452) was purchased from Thermo. Anti-Anti-NMNAT1 (ab45652, 30), PINK1 (ab23707, 66), MAVS (ab189303, 57), N-methyl-D-aspartate receptor (NMDAR) (ab134308, 105), CaMKI (ab68234, 41), TRAF6 (ab227560, 58), p-JNK (ab239886, 48) , JNK (ab208035, 48), DLK2 (ab171037, 45), p-MKK4 (ab52958, 44), MKK4 (ab33912, 44), MLK3 (ab51068, 93), Hippocalcin (ab24560, 23), SCG10 (ab115513, 21) , and ASC (ab155970) antibodies were purchased from Abcam. NAD/NADH Assay (ab65348), Calcium Assay Kit (ab102505), and Complex I Rodent Profiling ELISA Kit (ab136809) were obtained from Abcam. ROS detection kits (E004-1-1) were purchased from Nanjing Jiancheng Bioengineering Institute. Annexin V-FITC/PI kit (CA1020) was purchased from Solarbio.
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4

Evaluation of Danshen Bioactive Compounds

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DSS (NMR grade, ≥99.9%, Sigma-Aldrich, United States), sulfanilamide ≥99%, N-1-naphthylethelene (ACS reagent, ≥98%, Sigma Aldrich, United States), MTT ≥ 97.5% HPLC grade, LPS from Escherichia coli (Sigma Aldrich, United States).
Salvianolic acid A ≥ 98%, salvianolic acid B ≥ 95%, danshensu ≥ 98%, rosmarinic acid ≥ 98%, caffeic acid ≥ 98%, tanshinone IIA ≥ 98%, tanshinone I ≥ 96%, cryptotanshinone ≥ 98%, and dihydrotanshinone I ≥ 96% (Tauto Biotech, China).
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5

Aqueous Extract of T. sarmentosa

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The preparation of the aqueous extract of T. sarmentosa was described in a previous study [14 (link)]. The various components of the T. sarmentosa extract, including caffeic acid, rosmarinic acid, salvianolic acid A, and salvianolic acid B, were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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