Chemicals were from Sigma-Aldrich (St. Louis) unless otherwise noted. The flavonoids were purchased from Selleckchem (Houston, TX): resveratrol (Cat #S1396), fisetin (Cat #S2298), luteolin (Cat #S2320), rutin (Cat #S2350), epigallocatechin gallate (EGCG, Cat #S2250), curcumin (Cat #S1848), pirfenidone (Cat #S2907), and myricetin (Cat #S2326). Apigenin, catechin, and quercetin were purchased from Sigma-Aldrich (Cat #1760595, #1096790 and 1,592,409, respectively).
Epigallocatechin gallate (egcg)
Manufactured by Selleck Chemicals
Sourced in United States
EGCG is a pure polyphenol compound extracted from green tea leaves. It is a white crystalline powder with a molecular formula of C₂₂H₁₈O₁₁. EGCG is a natural antioxidant and its core function is to scavenge free radicals.
Automatically generated - may contain errors
Lab products found in correlation
Resveratrol, by Selleck Chemicals (2 mentions)
Rapamycin, by Selleck Chemicals (2 mentions)
Cb 839, by Selleck Chemicals (2 mentions)
Dimethyl α ketoglutarate, by Merck Group (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Gapdh, by Santa Cruz Biotechnology (2 mentions)
Rapamycin, by LC Laboratories (2 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions)
Fluo 4 am, by Thermo Fisher Scientific (2 mentions)
Fisetin, by Selleck Chemicals (1 mentions)
Pirfenidone, by Selleck Chemicals (1 mentions)
Luteolin, by Selleck Chemicals (1 mentions)
Apigenin, by Merck Group (1 mentions)
Curcumin, by Selleck Chemicals (1 mentions)
Rutin, by Selleck Chemicals (1 mentions)
Myricetin, by Selleck Chemicals (1 mentions)
Quercetin, by Merck Group (1 mentions)
Catechin, by Merck Group (1 mentions)
Disulfiram, by Bio-Techne (1 mentions)
Vincristine, by Selleck Chemicals (1 mentions)
17 protocols using epigallocatechin gallate (egcg)
1
Flavonoid Compounds Protocol
2
Giardicidal Activity and Cytotoxicity Assay
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The following inhibitors were purchased: Canavanine and metronidazole (Sigma Aldrich); vincristine, epigallocatechin gallate, and SNX-2112 (Selleck Chemicals); disulfiram (Tocris); and conoidin A (Cayman Chemical). For assays of giardicidal activity, serial 1:3 dilutions of these compounds were made in 96-well plates, G. lamblia WB or GS/M trophozoites were added, and cultures were grown for 1–2 days at 37°C under anaerobic conditions [29 (link)]. Parasite cell growth and viability were determined by measuring ATP levels with the BacTiter-Glo microbial cell viability assay reagent (Promega) in a microplate reader. The 50% effective concentration (EC50) was derived from the concentration-response curves using BioAssay software (Cambridge soft). Acute human cytotoxicity was assayed with the human epithelial cell line, HeLa (ATCC CCL-2) [29 (link),30 (link)]. Compounds were serially diluted (1:3) and added to HeLa cell cultures in 96-well plates. Cells were grown for two days, and viable cell numbers were determined using AlamarBlue reagent (Invitrogen). As done for the EC50 calculations, the 50% cytotoxic concentration (CC50) was derived from the normalized concentration-response curves using BioAssay software (Cambridge soft).
3
Apoptosis pathway inhibitors protocol
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ABT-263, A-1331852 and A-1210477 were from AbbVie (North Chicago, IL, USA),
ABT-199, epigallocatechin gallate (EGCG), CB-839, simvastatin, rapamycin and
torin-1 from Selleck Chemicals (Houston, TX, USA),
gamma-L-glutamyl-p-nitroanilide (GPNA) from Insight
Biotechnology (Wembley, Middlesex, UK), azaserine from Cambridge Bioscience
(Cambridge, UK), aminooxyacetate (AOA), sodium palmitate, dimethyl
α-ketoglutarate, oxaloacetate and citrate from Sigma-Aldrich
(Gillingham, UK), L-glutamine from Life Technologies (Paisley, UK) and
GSK2194069, SB204990, atorvastatin, pitavastatin and bafilomycin A1 from Tocris
(Abingdon, UK). Antibodies against PARP, BCL-2, MCL-1, BAX, BAK and GAPDH were
from Santa Cruz Biotechnology (Santa Cruz, CA, USA), caspase-3, caspase-9,
BCL-XL, BCL-w, BIM, PUMA, BAD, IDH2, ACL, ACO2, ATG5 and ATG7
from Cell Signaling Technology (MA, USA), BID from Prof. J. Borst (The
Netherlands Cancer Institute, Amsterdam, the Netherlands), NOXA from Millipore
(Watford, UK) and SLC1A5, GLS, GFAT, GLUD1, IDH3, FASN and HMGR from Abcam
(Cambridge, UK).
ABT-199, epigallocatechin gallate (EGCG), CB-839, simvastatin, rapamycin and
torin-1 from Selleck Chemicals (Houston, TX, USA),
gamma-L-glutamyl-p-nitroanilide (GPNA) from Insight
Biotechnology (Wembley, Middlesex, UK), azaserine from Cambridge Bioscience
(Cambridge, UK), aminooxyacetate (AOA), sodium palmitate, dimethyl
α-ketoglutarate, oxaloacetate and citrate from Sigma-Aldrich
(Gillingham, UK), L-glutamine from Life Technologies (Paisley, UK) and
GSK2194069, SB204990, atorvastatin, pitavastatin and bafilomycin A1 from Tocris
(Abingdon, UK). Antibodies against PARP, BCL-2, MCL-1, BAX, BAK and GAPDH were
from Santa Cruz Biotechnology (Santa Cruz, CA, USA), caspase-3, caspase-9,
BCL-XL, BCL-w, BIM, PUMA, BAD, IDH2, ACL, ACO2, ATG5 and ATG7
from Cell Signaling Technology (MA, USA), BID from Prof. J. Borst (The
Netherlands Cancer Institute, Amsterdam, the Netherlands), NOXA from Millipore
(Watford, UK) and SLC1A5, GLS, GFAT, GLUD1, IDH3, FASN and HMGR from Abcam
(Cambridge, UK).
4
Pharmacological Screening of Cellular Modulators
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Acetylcysteine, astaxanthin, epigallocatechin gallate, harmine, LY294002, metformin, moclobemide, phenformin, piperlongumine, rapamycin, resveratrol, sirtinol, spermidine, and tetrahydrocurcumin were purchased from Selleck Chemicals (Houston, TX, USA). rapamycin was purchased from LC Laboratories (Woburn, MA, USA); dihydroethidium, hydroxyurea, and phenformin hydrochloride were obtained from Sigma (St. Louis, MO, USA); and leptomycin B (LMB) was purchased from Alomone Labs (Jerusalem, Israel). AnnH75 was developed and synthesized in the Bracher Lab (Ludwig-Maximilians University, Munich, Germany) (16 (link)).
5
EGCG Treatment in Aged Mice
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Healthy and 3‐month‐, 16‐month‐ and 18‐month‐old SPF class c57bl/6 mice were purchased from the Experimental Animal Center in Chongqing Medical University (Chongqing, China). All procedures on experimental animals were approved by the Animal Care and Use Committee at the Chongqing Medical University. Animal experiments were performed in accordance with the NIH guidelines (Guide for the care and use of laboratory animals)13 . Sixteen‐month‐old mice were treated with single dose (50 mg/kg/day) of EGCG (Selleck) and dimethyl sulphoxide (DMSO, dissolvent control) by intraperitoneal injection for 8 weeks. The mice were killed by carbon dioxide asphyxia, and cardiac tissues were collected.
6
Natural Product Chemical Library Evaluation
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A chemical library composed of 800 single purified compounds derived from natural products were purchased from MicroSource Discovery Systems/Topscience (Shanghai, China). EGCG used in the rest experiments in this study was purchased from Selleck (Catalog No. S2250).
7
PC12 Cell Line Compound Evaluation
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The PC12 cell line was supplied by the Central Laboratory of the Central Hospital of Wuhan (Wuhan, China). The RPMI 1640 medium was purchased from Hyclone; GE Healthcare Life Sciences (Logan, UT, USA). Fetal bovine serum (FBS) was purchased from Zhejiang Tianhang Biotechnology Co., Ltd. (Hangzou, China). EGCG (>97.0%) was purchased from Selleck Chemicals (Houston, TX, USA). CORT (>97.0%) and the hedgehog-smoothened inhibitor cyclopamine (>99.0%) were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., (Shanghai, China). Desipramine (DIM; >98.0%), a well-known antidepressant (19 (link)) which was used as the positive control (18 (link)) in the present study, was purchased from Sigma Aldrich; Merck KGaA (Darmstadt, Germany). All pharmacological agents were prepared as a stock solution and stored at −20°C.
8
EGCG Ameliorates Abdominal Aortic Constriction
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Before surgery, the mice were randomly divided into 2 groups (30 mice in the AAC group and 15 mice in the sham group). After surgery, the AAC group was randomly divided into 2 groups (15 mice in the AAC + EGCG group and 15 mice in the AAC + vehicle group), and the sham group was classified as sham + vehicle group. On the fifth day postsurgery, 200 mg of EGCG (purity 99%, Selleck) was weighed and dissolved in 1 mL of dimethyl sulfoxide; this was diluted with 0.9% of stroke-physiological saline solution at a ratio of 1:20 and administered intraperitoneally at a rate of 50 mg/kg/d per mouse for 6 weeks. The sham + vehicle and AAC + vehicle groups were treated with the same amount of dimethyl sulfoxide diluted with 0.9% of stroke-physiological saline solution at a 1:20 ratio.
9
Cell Proliferation Assay with Glutamine and Enzyme Inhibitors
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Cell proliferation was determined by trypan blue exclusion assay (Thermo Fisher Scientific, Waltham, MA, USA). To investigate the effect of glutamine on cell growth, culture medium were supplemented with 2 mM L-glutamine (Gibco, 210-51-024), 4mM dimethyl-2-oxoglutarate (α-KG, Sigma-Alsdrich, 349631) and 2 mM nonessential amino acid (NEAA, Gibco, 11140), respectively. In addition, enzyme inhibitor epigallocatechin gallate (EGCG, selleckchem, S2250) at concentration of 0, 25, 50 and 100 μM and another enzyme inhibitor aminooxyacetate (AOA, selleckchem, S4989) at concentration of 0, 0.5, 1.0 and 2.0 mM were added to culture media, respectively, aiming to observe their effects on HCC cell growth.
For colony formation assay, cells were grown in puromycin-containing medium (1.0 μg/mL) for 10 days. Finally formed colonies were stained with 0.5% crystal violet.
For colony formation assay, cells were grown in puromycin-containing medium (1.0 μg/mL) for 10 days. Finally formed colonies were stained with 0.5% crystal violet.
10
Antibody Utilization for Cellular Analysis
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