As previously described, A549/A400 cells were cultured and grown in Dulbecco’s modified Eagle’s medium (DMEM) (GIBCO, 12100-046) supplemented with 10% foetal bovine serum (HyClone, SH30070.03) at 37 °C in a humidified atmosphere of 5% CO2. A549/A400 cells were generated and provided by Gwo-Tarng Sheu.15 (link) As previously described, the pemetrexed-resistant subline A549/A400 was established from parental A549 cells in a stepwise manner by exposure to increasing concentrations of pemetrexed. Briefly, A549 cells in low cellular density were seeded onto 10-cm Petri dish and treated with 50 nM pemetrexed until the surviving cells grew to an obvious colony. The selected colony was amplified in the presence of pemetrexed until confluence before the drug dose was increased in multiples of two for the next round of selection. As previously described, 3-Methyladenine (3-MA) (M 9281), chloroquine diphosphate salt (C 6628), and MG132 (474790) were purchased from Sigma (St. Louis, MO, USA). As previously described, GMI was provided by Mycomagic Biotechnology Co, Ltd. (Taipei, Taiwan), and GMI was generated and extracted as previously described.24 (link)
Chloroquine diphosphate salt c6628
Manufactured by Merck Group
Sourced in United States
Chloroquine diphosphate salt (C6628) is a chemical compound used in various laboratory applications. It is a salt form of the organic compound chloroquine. This product is typically utilized as a reagent or intermediate in scientific research and experiments, but its specific uses and applications can vary depending on the intended purpose of the research or analysis being conducted.
Automatically generated - may contain errors
Lab products found in correlation
Puromycin dihydrochloride, by Merck Group (2 mentions)
Polybrene tr 1003 g, by Merck Group (2 mentions)
Z vad ome fmk ab120487, by Abcam (2 mentions)
Leptomycin b l2913, by Merck Group (2 mentions)
N acetyl l cysteine a9165, by Merck Group (2 mentions)
Mg132, by Merck Group (1 mentions)
An3ca, by Thermo Fisher Scientific (1 mentions)
Hec 1a, by Merck Group (1 mentions)
Ishikawa, by Thermo Fisher Scientific (1 mentions)
Hec 1 a atcc htb 112, by American Type Culture Collection (1 mentions)
Hec59, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (1 mentions)
Pepstatin a 4397 v, by Peptide Institute (1 mentions)
Rapamycin tlrl rap, by InvivoGen (1 mentions)
E64d 4321 v, by Peptide Institute (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Fugene hd transfection reagent, by Promega (1 mentions)
Lipofactamine 2000, by Thermo Fisher Scientific (1 mentions)
Harmine, by Merck Group (1 mentions)
Hydrocortisone, by Thermo Fisher Scientific (1 mentions)
9 protocols using chloroquine diphosphate salt c6628
1
Pemetrexed-resistant A549 Cell Line Generation
2
Selective Inhibitors for Cell Research
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MG132 (ab141003), bortezomib (ab142123) and Z-VAD(OMe)-FMK (ab120487) were obtained from Abcam (Cambridge, UK). Chloroquine diphosphate salt (C6628), Cyclohexamide (C7698), Leptomycin B (L2913), N-Acetyl-L-cysteine (A9165), Doxycycline hyclate (D9891), G418 sulfate (345810), 12-O-Tetradecanoylphorbol-13-acetate or TPA (P1585), sodium butyrate (B5887), Polybrene (TR-1003-G) and Puromycin dihydrochloride (P8833) were brought from Sigma-Aldrich Corp. (St. Louis, MO, USA).
3
Endometrial Cancer Cell Lines Protocol
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AN3CA (ATCC, HTB-111) and HEC-1A (ATCC, HTB-112) cells were obtained from the American Type Culture Collection. HEC-59 (JCRB1120) and Ishikawa (JCRB1505) cells were purchased from the Japanese Collection of Research Bioresources (JCRB) cell bank. AN3CA cells were cultured and grown in Minimum Essential Medium (MEM) (GIBCO, 41500-034) supplemented with 10% fetal bovine serum (GIBCO, 10437). HEC-59 and Ishikawa cells were cultured in Minimum Essential Medium (MEM) (GIBCO, 41500-034) supplemented with 15% fetal bovine serum. HEC-1A cells were cultured in McCoy’s 5A Medium (SIGMA, M4892) supplemented with 10% fetal bovine serum. All cells were cultured at 37 °C in a humidified atmosphere of 5% CO2. PQR309 (Bimiralisib; 23441) and KJ-Pyr-9 (19116) were purchased from Cayman Chemical (Ann Arbor, MI). Chloroquine diphosphate salt (C 6628) was purchased from Sigma (St. Louis, MO, USA).
4
Huh7.5.1 Cell Culture and Antibodies for Autophagy
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KOD-plus- (KOD-201) was employed for high-fidelity polymerase chain reactions (TOYOBO). Huh7.5.1 cells derived from the Huh7 cell line (ATCC CCL-185) were cultured in a Dulbecco's modified Eagle Medium (DMEM; Wako, 045-30285) containing 10% fetal calf serum (JRH biosciences/SIGMA, 12603C) and 1% nonessential amino acids (Invitrogen, 11140050). Polyclonal antibodies against human LC3 was described previously [10] (link). The mouse monoclonal antibody, clone M2, against FLAG peptide (DYKDDDDK) (F1804), ammonium chloride (254134), and chloroquine diphosphate salt (C6628) were purchased from SIGMA-ALDRICH, the mouse monoclonal antibodies against GAPDH (ab8245) were from Abcam, and the rabbit monoclonal antibody against Myc epitope tag (2278) was from Cell Signaling Technology. Protein concentrations were determined using the bicinchoninic acid protein assay reagent (Pierce, 23225). E64d (4321-v) and pepstatin A (4397-v) were purchased from Peptide Institute, and the rapamycin (#tlrl-rap) was from InvivoGen. Torin1 was a kind gift from Dr. Nathanael S. Gray at the Dana Farber Cancer Institute and Dr. David Sabatini at the Whitehead Institute for Biomedical Research. To introduce the plasmid into the cells, FuGENE HD transfection reagent (E2311) was used (Promega).
5
Transfection and Treatment of HEK293 Cells
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HEK293 cells purchased from ATCC (NumberCRL-1573™) were cultured at 37°C in an incubator containing 5% CO2 as previously described [10 (link)]. Cells were transfected with lipofactamine2000 (11668–027; Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s guidelines. Lactacystin (426100) was purchased from Calbiochem (La Jolla, CA, USA). Chloroquinediphosphate salt (C6628), and harmine (286044) were purchased from Sigma-Aldrich (Louis, MO, USA).
6
Cell Culture Conditions for Breast Cancer Cell Lines
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MDA-MB-231, MDA-MB-231/sh-ATG5 (ATG5 knockdown [5 (link)]), MDA-MB-231/GFP-LC3 (stable GFP-LC3 overexpression [5 (link)]) and MCF-7 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) (Cellgro, 10-013-CV) containing fetal bovine serum (10%; Gibco, 26140), penicillin (100 U/ml) and streptomycin (100 μg/ml) (Gibco, 15240) at 37°C and 5% CO2 in a humidified incubator. T47D cells were cultured in RPMI 1640 medium under similar conditions. MCF-10A cells were maintained in DMEM/F12 media supplied with 5% horse serum, 20 ng/mL EGF, 0.5 mg/mL hydrocortisone, 100 ng/mL cholera toxin, 10 μg/mL insulin (Invitrogen, Carlsbad, CA, USA). Rapamycin (R8781), tamoxifen (T5648), bafilomycin A1 (B1793) and Chloroquine diphosphate salt (C6628) were from Sigma. Recombinant ADI-PEG20 was a gift of Polaris Pharmaceuticals Inc. (San Diego, CA).
7
Selective Inhibitors for Cell Research
8
Autophagy Inhibitors and Platinum-based Chemotherapy
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3-Methyladenine (3-MA) (S2767), Rapamycin (S1039), and Wortmannin (S2758) were purchased from Selleckchem (Houston, TX, USA). Chloroquine diphosphate salt (C6628) was purchased from Sigma-Aldrich (MO, USA). cis-Diamminedichloroplatinum (Cisplatin, DDP, Lot.No:8L012A88) was purchased from QILU Pharmaceutical (Shandong, China). Antibodies against ATG5 (ab109490), ATG7 (ab133528), MITF (ab140606), and LC3B (ab192890) were purchased from Abcam (Cambridge, UK). Antibodies against p62 (18420-1-AP) and GAPDH (60004-1-Ig) were purchased from Proteintech (Chicago, IL, USA). Antibody against LC3B (83506S) and Beclin 1 were purchased from Cell Signaling Technology (Danvers, MA, USA). LAMP-1 (sc-19992) and LAMP-2 (sc-18822) were purchased from Santa Cruz (Dallas, TX, USA).
9
Autophagy and Epithelial-Mesenchymal Transition
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Antibodies against SQSTM1 (PB0458, 1:400) was obtained from Boster Biological Technology Co. Ltd. Antibody against MFN1 (ab107129), LC3B (ab48394), Pink (ab23707), Parkin (ab77924) and Snail (ab53519) were purchased from Abcam. Antibodies against BECN-1 (sc-48341) and Fis 1(sc-376469) were purchased from Santa Cruz Biotechnology, Inc. Antibodies against N-cadherin (#13116) and E-cadherin (#14472) were obtained from Cell Signalling Technology. The Cell Counting Kit-8 kit (C0037) was provided by Beyotime Institute of Biotechnology. Bromodeoxyuridine (BrdU) proliferation assay kit (2750) was purchased from Millipore Corporation. The immunocytochemistry detection kits (SPN-9001) obtained from ZSGB-BIO. Chloroquine diphosphate salt (C6628) was purchased Sigma. mRFP-eGFP-LC3 plasmid was obtained from Hanbio Biotechnology Co. Ltd (Shanghai, China). Enhanced chemiluminescence (ECL, RPN2236) reagents were from Amersham International. All other reagents were from common commercial sources.
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