Hepa1-6 cells (hepatocellular carcinoma cell line) were obtained from Wuhan University. The cells were cultured in Dulbecco’s modified Eagle medium (Servicebio, Wuhan, China) with 1% penicillin-streptomycin (Biosharp, Hefei, China) and 10% fetal bovine serum (Gibco, Grand Island, NY, United States) at 37 °C with 5% CO2. The Candida albicans SC5314 standard strain was purchased from Biofeng Lab (Shanghai, China) and inoculated in yeast extract-peptone-D-glucose (YPD, Hopebio, Qingdao, China) and cultured for 24 h with continuous shaking at 30 °C overnight. The Malassezia furfur ATCC14521 strain was purchased from Fenghui Biotechnology Co., Ltd (Changsha, China) and inoculated in 2693 Modfied Dxn (mDxan, Hopebio, Qingdao, China) and cultured for 48 h with continuous shaking at 30 °C overnight.
Dulbecco s modified eagle medium
Manufactured by Wuhan Servicebio Technology
Sourced in China
Dulbecco's modified Eagle medium (DMEM) is a cell culture medium commonly used for the cultivation of various cell types. It provides a balanced salt solution with a variety of nutrients, including amino acids, vitamins, and other essential components to support cell growth and proliferation in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Biosharp (1 mentions)
Trypsin, by Wuhan Servicebio Technology (1 mentions)
Tomm20, by ABclonal (1 mentions)
P pi3k, by ABclonal (1 mentions)
P akt, by ABclonal (1 mentions)
Cell counting kit 8 (cck8), by Sparkjade (1 mentions)
Beclin1, by ABclonal (1 mentions)
β actin, by ABclonal (1 mentions)
Fetal bovine serum (fbs), by Procell (1 mentions)
Penicillin streptomycin, by Wuhan Servicebio Technology (1 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Gentamycin, by Sangon (1 mentions)
Amikacin, by Solarbio (1 mentions)
Raw264, by American Type Culture Collection (1 mentions)
Triton x 100, by Sangon (1 mentions)
6 protocols using dulbecco s modified eagle medium
1
Establishing Hepatocellular Carcinoma and Fungal Cell Models
2
Investigating Mechanisms of Cell Death
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UP302 was purchased from Nanjing m&m biotechnology Co. Ltd. The following reagents were used: fetal bovine serum (Procell, China), Dulbecco's Modified Eagle Medium (DMEM), Iscove's Modified Dulbecco's Medium(IMDM), trypsin (Servicebio, China), CCK8 (Sparkjade, China), chloroquine (Sparkjade, China), PI and FITC, Mitochondrial membrane potential assay kit with JC-1 and Reactive Oxygen Species Assay Kit (Beyotime, China), Antibodies against Caspase-3, BAX, BCL-2, LC3B, AKT123, PI3K, P-PI3K, P-AKT, AIF, Tomm20, PINK1, Parkin, Beclin1, and β-actin were purchased from ABclonal(China). Twelve Balb/c female nude mice, aged four weeks, were purchased from Jinan Pengyue Experimental Animal Breeding Co. LTD.
3
Microglia BV2 Cell Culture and Ruxolitinib Treatment
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The microglia cell line BV2 (Cat# 0356; RRID: CVCL_0182), which has been short tandem repeat genotyped, was purchased from the Institute of Cell Research, Chinese Academy of Medical Sciences (Shanghai, China). Cells were cultured in Dulbecco's modified Eagle medium (Cat# G4510; Servicebio, Wuhan, China) with 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and 1% penicillin-streptomycin (Servicebio), as previously described (Hong et al., 2020). RUX (Cat# ab141356; Abcam, Cambridge, MA, USA) was dissolved in pH 3.5 citrate buffer according to a previous study (Das et al., 2016). Microglia were pretreated with 0.1, 0.5, or 1 mM RUX for 24 hours, followed by treatment with IFN-γ (20 ng/mL; Invitrogen) for 6 hours.
4
Culturing Duck Embryo Cells with NDRV Strain
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DEFs obtained from 11-day-old duck embryos were cultured in high glucose Dulbecco's modified Eagle medium (DMEM; Servicebio, Wuhan, China) supplemented with 10% fetal bovine serum (FBS) in a cell incubator at 37°C. The NDRV N20 strain used in this work was obtained from the veterinary molecular etiology laboratory of Shandong Agricultural University, which was verified as an attenuated strain by pathogenicity experiments (Yan et al., 2021 (link)).
5
Evaluating SWT's Anti-Ferroptosis Effect
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Mice liver parenchymal cell line AML12 was obtained from ATCC and cultured under the atmosphere of 5% CO 2 at 37 °C. The cells were cultured in Dulbecco's modified Eagle medium (Servicebio) supplemented with 10% fetal bovine serum (Gibco), penicillin G (100 U/mL) and streptomycin (100 μg/mL). To mimic the damage of hepatitis with ferroptosis, AML12 cells were exposed to a combination of OA and PA (500: 250 μM), as well as erastin (10 μM), for 24 h. Concurrently, different concentrations of SWT (25, 50, 100 mg/mL) or ferrostatin-1 (Fer-1, 1 μM) were administered to the cells for 24 h to evaluate the anti-ferroptosis effect of SWT. Subsequently, the cells were collected for further experimentation.
6
Phagocytosis Assay of Macrophages
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The phagocytosis assay was performed using the mouse macrophage cell line RAW 264.7 (ATCC catalogue number TIB-71) and peritoneal macrophages as previously described53 . Peritoneal macrophages were collected from 7-week-old female C57BL/6J mice on the 5th day after intraperitoneal injection of 1 ml 5% thioglycollate broth (BD Difco, catalogue number 211716)54 . Briefly, 5 × 105 cells were infected with bacteria at a multiplicity of infection of 50 for 1 h. The cells were then washed three times with 1× PBS and treated with 500 μl Dulbecco’s modified eagle medium (Servicebio, catalogue number G4515) containing 300 μg ml−1 gentamycin (Sangon Biotech, catalogue number A506614) or amikacin (Solarbio, catalogue number A9660) for 1 h to eliminate the extracellular bacteria. After washing three times with 1× PBS, the cells were treated with 1 ml of 1% TritonX-100 (Sangon Biotech, catalogue number A110694) for 10 min to release the intracellular bacteria. Then intracellular bacteria were counted by serial dilution agar plating method.
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