Cells were cultured on glass cover slides and treated with either different concentrations of TSN (0, 60, 80 and 100 nmol/l) or transfected with pcDNA3.1(+) (2 µg/ml), pcDNA3.1(+)-miR-23a-3p (2 µg/ml), miR-23a-3p-I-NC (1.0×10−4 mmol/l) or miR-23a-3p-I (1.0×10−4 mmol/l) for 48 h. Subsequently, cells were rinsed twice with PBS and fixed with 4% paraformaldehyde in PBS for 10 min. Next, the cells were stained with 0.1 mg/ml acridine orange (Beijing Dingguo Changsheng Biotechnology Co., Ltd.) for 1–2 min. The images were captured using a Leica TCS SP8 confocal laser-scanning microscope (Leica Microsystems GmbH) at 488 nm excitation and 515 nm emission wavelengths (magnification, ×63).
Acridine orange
Manufactured by Dingguo
Sourced in China
Acridine orange is a fluorescent dye used in various laboratory applications. It is a versatile stain that can bind to both DNA and RNA, emitting different colored fluorescence upon excitation. This property makes it a useful tool for various biological and biochemical analyses.
Automatically generated - may contain errors
Lab products found in correlation
Tcs sp8 confocal laser scanning microscope, by Leica (1 mentions)
Cell cycle and apoptosis detection kit, by Beyotime (1 mentions)
Ethidium bromide, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Mitochondrial membrane potential assay kit with jc 1, by Beyotime (1 mentions)
Chlorpromazine, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Monensin, by Merck Group (1 mentions)
Mk571, by Merck Group (1 mentions)
Lyso tracker red dye, by Beyotime (1 mentions)
Rpmi 1640 growth medium, by Thermo Fisher Scientific (1 mentions)
Verapamil vrp, by Merck Group (1 mentions)
Pamam g4 nh2, by Merck Group (1 mentions)
Pamam g4 oh, by Merck Group (1 mentions)
Fumitremorgin c, by Merck Group (1 mentions)
Fluorescein isothiocyanate (fitc), by Merck Group (1 mentions)
2 deoxyglucose, by Merck Group (1 mentions)
Brefeldin a, by Merck Group (1 mentions)
Bafilomycin a1, by Merck Group (1 mentions)
4 protocols using acridine orange
1
Acridine Orange Staining of TSN-Treated Cells
2
Mitochondrial Function and Cell Cycle Assays
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The mitochondrial membrane potential assay kit with JC-1 (catalog no. C2006), and the cell cycle and apoptosis detection kit (catalog no. C1052) were purchased from Beyotime Institute of Biotechnology (Haimen, China). Ethidium bromide (catalog no. E8751), dimethyl sulfoxide (DMSO) and MTT were purchased from Sigma-Aldrich (EMD Millipore, Billerica, MA, USA). Acridine orange (catalog no. 1AB10220) was purchased from Ding Guo Changsheng Biotechnology Co., Ltd. (Beijing, China). The novel tumstatin was designed by Laboratory of Chemistry and Molecular Biology Institute of Frontier Medical Sciences, Jilin University (Changchun, China) and synthesized by GL Biochem Ltd. (Shanghai, China), with a purity of 98%. Carmustine (catalog no. 1407011) was purchased from Jinyao Pharmaceutical Co., Ltd. (Tianjin, China; serial number).
3
Evaluation of Endocytosis Inhibitors
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PAMAM-G4-NH2, PAMAM-G4-OH, FITC, verapamil (VRP), fumitremorgin C (FTC), MK571, chlorpromazine, sodium azide, 2-deoxyglucose, ammonium chloride, brefeldin A, monensin, and bafilomycin A1 were purchased from Sigma-Aldrich Co. (St Louis, MO, USA). Dulbecco’s Modified Eagle’s Medium (DMEM) growth medium, Roswell Park Memorial Institute 1640 (RPMI-1640) growth medium, fetal bovine serum, and penicillin–streptomycin were purchased from Gibco BRL (Gaithersberg, MD, USA). Acridine Orange (AO) was purchased from DingGuo Biotechnology Co., Ltd. (Beijing, People’s Republic of China). Hoechst 33258 and LysoTracker Red dye were purchased from Beyotime Biotechnology Co., Ltd. (Nantong, People’s Republic of China). All other chemicals were commercially available reagents of at least analytical grade.
4
Investigating MAPK Signaling Pathways
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MAPK Family Antibody Sampler Kit (#9926) and Phospho-MAPK Family Antibody Sampler Kit (#9910) were purchased from Cell Signaling Technology (Beverly, MA). Anti-CEP-1 (cC-18) (sc-135460) antibody was obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Paraquat (PQ), Cyclosporin A (CsA), Bongkrekic acid (BA) and 4,4′-Diisothiocyanatostilbene-2,2′-disulfonic acid disodium salt hydrate (DIDS) were from Sigma. N-acety-L-cysteine (NAC) was purchased from TCI (Shanghai, China). Total Antioxidant Capacity Assay Kit with the ABTS Method (S0119), Fluo-3 AM (S1056), Hoechst 33342 (C1022), Mitochondrial Membrane Potential Assay Kit with JC-1 (C2006), Enhanced BCA Protein Assay Kit (P0010S) and Superoxide dismutase (SOD, S0088) were obtained from Beyotime (Shanghai, China). Acridine orange (AO) was from Dingguo Changsheng Biotechnology (Beijing, China). H2DCF-DA (D399), MitoSox (M36008), Mitotracker red (M7512), ATP determination kit (A22066) and SYTO 12 (S7574) were purchased from Molecular Probes (Eugene, Oregon, USA). PVDF membranes and ECL plus detection kit were obtained from Millipore (Bedford, MA, USA).
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