Rat Schwann Cell Line (RSC96 cells) were kindly provided by the Cell Bank of China. Cells were cultured in D-MEM culture medium supplemented with 10% (v/v) fetal bovine serum and 100U/mL penicillin & streptomycin. For treatment of H2O2, cells were firstly seeded in 6-plate wells at a density of approximately 50% at the second day, then, different dozes of H2O2 was added into the culture medium and stimulated for indicated times. Unless otherwise noted, the concentrations for H2O2-induced RSC96 cell injury were selected at 600 μM, and treated for 8 hrs. The ROS inhibitor, N-Acetyl-L-cysteine (NAC, HY-B0215, MedChemExpress), were utilized at the final concentrations of 10μM in this study. For the inhibition of RIPK1, 500 nM Necrostatin-1 (Nec-1, HY-15760, MedChemExpress) were added prior to H2O2 stimulation.
Hy 15760
Manufactured by MedChemExpress
Sourced in United States
HY-15760 is a benchtop centrifuge used for the separation and isolation of various biological samples such as cells, proteins, and nucleic acids. It provides high-speed centrifugation capabilities to facilitate common laboratory procedures.
Automatically generated - may contain errors
Lab products found in correlation
Hy 100579, by MedChemExpress (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Hy b0215, by MedChemExpress (1 mentions)
D2650, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Ncm460, by American Type Culture Collection (1 mentions)
Ccl 228, by American Type Culture Collection (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Sw480, by American Type Culture Collection (1 mentions)
Ab8978, by Abcam (1 mentions)
Ab202445, by Abcam (1 mentions)
Ab236868, by Abcam (1 mentions)
Ab6789, by Abcam (1 mentions)
Ab6721, by Abcam (1 mentions)
Hy 17589a, by MedChemExpress (1 mentions)
Irdye 800cw goat anti rabbit secondary antibody, by LI COR (1 mentions)
A20798, by ABclonal (1 mentions)
Hy 10431, by MedChemExpress (1 mentions)
Rm02821, by ABclonal (1 mentions)
A5243, by ABclonal (1 mentions)
6 protocols using hy 15760
1
Modeling Oxidative Stress in Rat Schwann Cells
2
Pharmacological Intervention in Rat Model
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Nec-1 (MedChem Express, HY-15760), zVAD-fmk (MedChem Express, HY-16658), and 3-MA (MedChem Express, HY-19312) were dissolved in 0.1% dimethylsulfoxide (DMSO, Sigma Aldrich, D2650) with 0.1 M phosphate-buffered saline (PBS). The drug dosages are: Nec-1 group, 0.6 mg/kg/day; zVAD-fmk group, 1.0 mg/kg/day; 3-MA group, 15 mg/kg/day; and DMSO group, 0.1% DMSO. Rats were injected intraperitoneally on day 0 and day 8, respectively. On day 21, rats were sacrificed after the first injection.
3
Plumbagin's Cytotoxic Effects on Colon Cancer
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The normal colonic epithelial cell line NCM460 (CL-0393) and the colon cancer cell lines SW480 (wild-type p53, CCL-228) and HCT116 (wild-type p53, CCL-247 EMT) were purchased from ATCC (USA). The cells were all cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum (FBS) and 100 × penicillin-streptomycin solution in an incubator at 37 °C with 5% CO2 and 95% humidity.
The cells were treated with plumbagin (99%, CFN90444; Wuhan ChemFaces Biochemical Co., Ltd., Wuhan, China) at final concentrations of 0, 5, 10, 15 and 20 μmol/L for 24, 48 and 72 h. Thereafter, plumbagin was used at a concentration of 15 μmol/L for subsequent experiments (72 h). To assess the safety of plumbagin, normal colonic epithelial cells (NCM460) were treated with 15-μmol/L plumbagin for 72 h. The resulting cell viability was 72.6 ± 7.1%, indicating that the dosage was relatively safe.
Before plumbagin treatment, the cells underwent treatment with the ferroptosis inhibitors Fer-1 (1 μmol/L, HY-100579, MedChemExpress) and Lip-1 (0.2 μmol/L, HY-132216) for 24 h. Additionally, the cells were pre-treated with 10 μmol/L Nec-1 (HY-15760) and 10 μmol/L Z-VAD-FMK (HY–16658B) for 24 h to inhibit necroptosis and pan-caspase apoptosis, respectively.
The cells were treated with plumbagin (99%, CFN90444; Wuhan ChemFaces Biochemical Co., Ltd., Wuhan, China) at final concentrations of 0, 5, 10, 15 and 20 μmol/L for 24, 48 and 72 h. Thereafter, plumbagin was used at a concentration of 15 μmol/L for subsequent experiments (72 h). To assess the safety of plumbagin, normal colonic epithelial cells (NCM460) were treated with 15-μmol/L plumbagin for 72 h. The resulting cell viability was 72.6 ± 7.1%, indicating that the dosage was relatively safe.
Before plumbagin treatment, the cells underwent treatment with the ferroptosis inhibitors Fer-1 (1 μmol/L, HY-100579, MedChemExpress) and Lip-1 (0.2 μmol/L, HY-132216) for 24 h. Additionally, the cells were pre-treated with 10 μmol/L Nec-1 (HY-15760) and 10 μmol/L Z-VAD-FMK (HY–16658B) for 24 h to inhibit necroptosis and pan-caspase apoptosis, respectively.
4
Probing Ferroptosis and EMT Mechanisms
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RSL3 (HY-100218, MedChemExpress), chloroquine (CQ) (HY-17589A, MedChemExpress), necrostatin-1 (Nec-1) (HY-15760, MedChemExpress), ferrostatin-1 (Fer-1) (HY-100579, MedChemExpress), SB431542 (HY-10431, MedChemExpress), and carbon 11 (C11)-BODIPY581/591 (RM02821, Abclonal, Wuhan, China) were the reagents used.
Nodal (A9902, Abclonal; sc-373910, Santa Cruz Biotechnology), SCD1 (ab236868, Abcam), Phospho-Smad2 (Ser465/467)/Smad3 (Ser423/425) (8828 S, Cell Signalling Technology), Smad2/3 (ab202445, Abcam; 5678, Cell Signalling Technology), E-cadherin (A20798, Abclonal), N-cadherin (A0432, Abclonal), vimentin (ab8978, Abcam), snail (A5243, Abclonal), GAPDH (A19056, Abclonal), β-tubulin (AC008, Abclonal), goat anti-rabbit immunoglobulin (Ig) G heavy and light chain (H&L) (horse radish peroxidase [HRP]) (ab6721, Abcam), goat anti-mouse IgG (H&L) (HRP) (ab6789, Abcam), IRDye 680 donkey anti-mouse IgG-(H + L)/goat anti-rabbit IRDye 800CW secondary antibody (926-68072/926-32211, LI-COR Biosciences, Lincoln, NE, USA) were the antibodies used in this study.
Nodal (A9902, Abclonal; sc-373910, Santa Cruz Biotechnology), SCD1 (ab236868, Abcam), Phospho-Smad2 (Ser465/467)/Smad3 (Ser423/425) (8828 S, Cell Signalling Technology), Smad2/3 (ab202445, Abcam; 5678, Cell Signalling Technology), E-cadherin (A20798, Abclonal), N-cadherin (A0432, Abclonal), vimentin (ab8978, Abcam), snail (A5243, Abclonal), GAPDH (A19056, Abclonal), β-tubulin (AC008, Abclonal), goat anti-rabbit immunoglobulin (Ig) G heavy and light chain (H&L) (horse radish peroxidase [HRP]) (ab6721, Abcam), goat anti-mouse IgG (H&L) (HRP) (ab6789, Abcam), IRDye 680 donkey anti-mouse IgG-(H + L)/goat anti-rabbit IRDye 800CW secondary antibody (926-68072/926-32211, LI-COR Biosciences, Lincoln, NE, USA) were the antibodies used in this study.
5
Doxorubicin-Induced Cardiotoxicity Assay
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Embryos were obtained through natural spawning. Tg(cmlc2: eGFP) fish larvae of 30 hpf were arrayed into 24‐well plates(5 fishes per well) and were treated with Dox(65 µM; S1208, Selleck Chemicals, USA) in the presence of FeCl3(10 µM; 451649, Sigma‐Aldrich, USA) for 30 h. The embryo medium was then replaced with fresh E3 buffer (without Dox) and continued to culture for 40 h. At 100 hpf, fluorescent heart‐beating images were acquired by Leica DMI 3000B inversed microscope system (Leica Microsystems, Germany). For the DXZ‐treated group, 450 µM DXZ (S1222, Selleck Chemicals, USA) was supplemented in the embryo medium during and after Dox treatment from 30hpf to 100hpf. For the test with different cell death inhibitors, Fer‐1 (1 µm; HY‐100579, MedChemExpress, USA), 3‐MA (250 µM; HY‐19312, MedChemExpress, USA), Emricasan (50 µM; HY‐10396, MedChemExpress, USA), Nec‐1 (10 µM; HY‐15760, MedChemExpress, USA) were administered from 30hpf to 100 hpf.
6
Cell Line Maintenance and Treatment
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The human HCC cell lines (Huh7, HepG2) were obtained from the cell bank of the Type Culture Collection of the Chinese Academy of Sciences. HEK-293T and HCC cell lines (SM-386, YY-8103, MHCC97-H) were obtained from ATCC cell bank, and routinely maintained in Dulbecco’s modified Eagle’s medium (DMEM, Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco). L02 cells were purchased from Shanghai Zhong Qiao Xin Zhou Biotechnology Co. Ltd., and cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco) supplemented with 10% FBS. All cells were maintained in a humidified 5% CO2 atmosphere at 37 °C, and tested for mycoplasma contamination every month. FCN3-overexpressed MHCC97-H, YY-8103 and control cells were treated with T0901317 (HY-10626, MedChemExpress), Erastin (HY-15736, MedChemExpress), FINO2 (HY-129457, MedChemExpress), AKTi-1/2 (HY-10355, MedChemExpress), Ferrostatin-1 (SML0583, Sigma-Aldrich), N-acetyl-L-cysteine (A9165, Sigma-Aldrich), Z-VAD-fmk (FMK001, R&D Systems), and Necrostatin-1 (HY-15760, MedChemExpress) for indicated time.
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