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2 protocols using neutral red solution nr

1

Cellular Assays for Oxidative and Nitrosative Stress

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Dulbecco's Modified Eagle Medium (DMEM), Fetal Bovine Serum (FBS), and penicillin/streptomycin were purchased from Gibco (Grand Island, USA). Fluoromount G and DAPI were obtained from Life Technologies (Carlsbad, USA). LysoTracker probe, 2′, 7′-dichlorofluorescein diacetate (DCFH-DA) reactive oxygen species (ROS)-sensitive fluorescent dye, and the NO specific fluorescent probe DAF-FM Diacetate were commercially obtained (Thermo Scientific, Massachusetts, USA). [4, 5-dimethyl-thiazol-2-yl] -2, 5-diphenyltetrazolium bromide (MTT), neutral red solution (NR), and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, USA). Agalsidase-β and chloroquine were purchased from Genzyme (Boston, USA) and Cristália (São Paulo, Brazil), respectively. All other reagents were obtained from Sigma-Aldrich (St. Louis, USA) if not otherwise specified.
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2

Lysosomal Dynamics in Cell Culture

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RPMI 1640 medium (RPMI), fetal bovine serum (FBS), collagen I from rat tail, and Penicillin/Streptomycin were purchased from Gibco (Grand Island, NE, USA). Fluoromount G and DAPI were obtained from Life Technologies (Carlsbad, CA, USA). LysoTracker® DND-99 probe, Alexa Fluor® 647 Phalloidin, and Actin Red™555 ReadyProbes™ Rhodamine phalloidin were commercially obtained (Thermo Scientific, Waltham, MA, USA). Murine Interferon Gamma (IFN-γ) was purchased from Prepotech. [4, 5-dimethyl-thiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT), neutral red solution (NR), and dimethyl sulfoxide (DMSO) were obtained from Sigma-Aldrich (St. Louis, MO, USA). CQ was purchased from Cristália (São Paulo, Brazil). Western Blotting reagents were obtained from GE HealthCare Life Sciences (Little Chalfont, UK). All other reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA) if not otherwise specified.
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