Pazopanib HCl (GW786034 HCl), salubrinal, and N‐acetylcysteine were purchased from Selleckchem. Cell counting kit‐8 (CCK‐8) was purchased from Beyotime. Dimethyl sulfoxide (DMSO), hydroxypropylmethylcellulose, and Tween‐80 were obtained from Sigma‐Aldrich. The RPMI‐1640 medium, fetal bovine serum (FBS), and penicillin/streptomycin were purchased from Biological Industries. All other reagents were purchased from Sigma‐Aldrich.
N acetylcysteine
Manufactured by Selleck Chemicals
Sourced in United States
N-acetylcysteine is a chemical compound used in various laboratory applications. It is a synthetic derivative of the amino acid cysteine and serves as a precursor to the antioxidant glutathione. N-acetylcysteine is commonly utilized in analytical and research settings due to its chemical properties.
Automatically generated - may contain errors
Lab products found in correlation
Hydroxypropyl methylcellulose, by Merck Group (1 mentions)
Rpmi 1640 medium, by Sartorius (1 mentions)
Penicillin streptomycin, by Sartorius (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Tween 80, by Merck Group (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
L arginine, by Merck Group (1 mentions)
C11 bodipy 581 591, by GLPBIO (1 mentions)
Pri 724, by Selleck Chemicals (1 mentions)
Mouse igg1, by Cell Signaling Technology (1 mentions)
Kg 501, by Bio-Techne (1 mentions)
Urolithin a, by Selleck Chemicals (1 mentions)
Z vad fmk, by Selleck Chemicals (1 mentions)
Mg132, by Selleck Chemicals (1 mentions)
Doxorubicin, by Selleck Chemicals (1 mentions)
Ara c, by Selleck Chemicals (1 mentions)
Fascin, by Santa Cruz Biotechnology (1 mentions)
Ab175186, by Abcam (1 mentions)
Ab125066, by Abcam (1 mentions)
10 protocols using n acetylcysteine
1
Cytotoxic Effects of Pazopanib, Salubrinal, and N-Acetylcysteine
2
Induction of Acute Pancreatitis via Pharmacological Agents
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J2 (GC38503), DHE (GC30025), and C11 BODIPY 581/591 (GC40165) were purchased from Glpbio, America. Erastin (S7242), caerulein (C9026) and N-acetylcysteine (S1623) were purchased from Selleck Chemicals, Japan. Soybean trypsin inhibitor (T6522), taurocholate acid (T4009), and L-arginine (A5131) were purchased from Sigma‒Aldrich, America. J2 (25 mg/kg) was administered via intraperitoneal injection for three consecutive days before establishing the AP model.
3
Modulation of Type I Interferon Signaling
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Anti–mouse α-IFNAR was purchased from Leinco and was used at concentrations ranging from 0.1 μg/mL to 10 μg/mL, as indicated (I-401, clone MAR1-5A3). Mouse isotype control antibody (mouse IgG1, Cell Signaling Technology, 5415) was used as a negative control for this antibody where applicable. Nec-1 was used to inhibit necroptosis; it was purchased from Selleckchem and used at 50 μM, unless otherwise specified (catalog S8037). Pan-caspase inhibitor Z-VAD-FMK was used at 10 μM to block apoptosis and was purchased from Selleckchem (catalog S7023). MG132 was used at 10 μM to inhibit proteasomal degradation of proteins and was purchased from Selleckchem (catalog S2619). Pharmacological inhibitors of the CREB/CBP/β-catenin system were purchased from Selleckchem (PRI-724, catalog S8262; KG-501, catalog S8409; and C646, catalog S7152) and Tocris Biosciences (666-15, catalog 5661). Purified mouse IFN-β was purchased from PBL Assay Science (catalog 12405-1). Urolithin A was purchased from Selleckchem (catalog S5312) and was used as an inducer of mitophagy. N-acetylcysteine was purchased from Selleckchem (catalog S1623) and used as an antioxidant.
4
Cancer Treatment Protocol Evaluation
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Ara-C (cytarabine), Doxorubicin (DOX), N-acetylcysteine (NAC) and Tert-butylhydroquinone (tBHQ) were all purchased from Selleck Chemicals (Selleck, USA).
5
Antibodies and Chemicals in Ferroptosis Assay
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The following antibodies were used in the present study at the indicated dilution for WB analysis, IP, IHC: Fascin (sc-21743, Santa Cruz; 1:1000 for WB analysis, 2 µg per 500 µg of total protein for IP, and 1:400 for IHC), xCT (12691S, CST; 1:1000 for WB analysis and 1:50 for IP), xCT (ab175186, Abcam; 1:200 for IHC), 4HNE (ab46545, Abcam; 1:200 for IHC), E-cadherin (3195S, CST;1:1000 for WB analysis), N-cadherin (13116S, CST;1:1000 for WB analysis), Vimentin (5741S, CST;1:1000 for WB analysis), ZEB1 (3396S, CST;1:1000 for WB analysis), Snail (3879S, CST;1:1000 for WB analysis), Slug (9585S, CST;1:1000 for WB analysis), GPX4 (ab125066, Abcam; 1:1000 for WB analysis), TFRC (ab214039, Abcam; 1:1000 for WB analysis), ACSL4 (sc-271800, Santa Cruz; 1:1000 for WB analysis), ub (sc-8071, Santa Cruz; 1:500 for WB analysis), β-actin (sc-47778, Santa Cruz; 1:2000 for WB analysis), and GAPDH (sc-47724, Santa Cruz; 1:2000 for WB analysis).
The chemicals erastin (S7242), N-acetylcysteine (S1623), ferrostatin-1 (S7243), liproxstatin-1 (S7699), Z-VAD-FMK (S7023), MG132 (S2619), cycloheximide (CHX) (S7418) and chloroquine (CQ) (S6999) were purchased from Selleck. 3-Methyladenine (3-MA) (189490) was obtained from Sigma Aldrich, and NP-G2-044 (HY-125506) was purchased from MedChemExpress (MCE).
The chemicals erastin (S7242), N-acetylcysteine (S1623), ferrostatin-1 (S7243), liproxstatin-1 (S7699), Z-VAD-FMK (S7023), MG132 (S2619), cycloheximide (CHX) (S7418) and chloroquine (CQ) (S6999) were purchased from Selleck. 3-Methyladenine (3-MA) (189490) was obtained from Sigma Aldrich, and NP-G2-044 (HY-125506) was purchased from MedChemExpress (MCE).
6
Mitophagy Assay with Oxidative Stress
7
Mouse Leydig Cell Isolation and Culture
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Mouse MLTC-1 Leydig cell line was obtained from ATCC (Manassas, VA, USA) and cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS) at 37°C, in a humidified 5% CO2. For primary mouse Leydig cell culture, male adult CD-1 mice were sacrificed, and the testes were washed in phosphate-buffered saline (PBS) and digested with 0.5% (w/v) collagenase IV (Sigma, St. Louis, MO, USA) for 30 min at 37°C. The reaction was stopped with addition of FBS and the supernatant was filtered with 400 mesh stainless screen. Collected cells were suspended in DMEM culture medium supplemented with 10% FBS, and incubated at 37°C under 5% CO2. Testosterone levels were measured with an ELISA kit (Jiancheng Bioengineering, Nanjing, China), according to the manufacturer’s instructions. Cell viability was determined using a Cell Proliferation Reagent Kit (MTT; Beytime, Nantong, China). ROS and ATP assay kits were purchased from Beytime (Nantong, China). PTC-209 and N-acetylcysteine (NAC) were obtained from Selleck (Houston, TX, USA) and Sigma (St. Louis, MO, USA), respectively. P16 siRNA (5ʹ- UUAUGUAUUUUUUAAAGCCAC-3ʹ) and P19 siRNA (5ʹ- AACUCUAUGAUCAUUUGCCGG-3ʹ) were synthesized from genepharma (Shanghai, China).
8
N-acetylcysteine Modulates Cellular Pathways
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N-acetylcysteine was obtained from Selleck (S1623). Cell Signaling Technology (Danvers, MA, USA) provided antibodies against FOXO3a (#2497), phospho-FOXO3a (#13129), BCL2L11 (#2819), CDKN1A (#2947), CDKN1B (#3686), and c-Myc (#5605). Affinity Biosciences provided the anti-GAPDH antibody (AF7021). Abcam (Cambridge, MA, USA) provided anti-FLAG (ab205606), anti-U2AF1 (ab197591), anti-NLRP3 (ab210491), anti-Caspase-1 (ab207802), anti-ASC (ab151700), anti-AKT (ab32505), anti-phospho-AKT (ab192623), anti-Bcl-2 (ab182858), anti-Bax (ab32503), and anti-Cleaved Caspase-3 (ab32042) antibodies. Autophagy Antibody Sampler Kit (#4445) was obtained from Cell Signaling Technology (Danvers, MA, USA). Sangon Biotech (Shanghai, China) provided HRP-conjugated secondary antibodies.
9
Intracellular ROS and Singlet Oxygen Detection
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Intracellular production of ROS was determined using Reactive Oxygen Species Assay Kit (Beyotime, S0033). After 24 h incubation with different treatment, cells were incubated with 10 μM 2′,7′-dichlorodihydrofluorescein diacetate dye (DCFH-DA) in DMEM for 30 min. Then, cells were washed with PBS, trypsinized and resuspended in PBS. The green fluorescence (DCFH-DA), corresponding to ROS levels, was determined using flow cytometer, microplate reader and fluorescence microscope. N-acetylcysteine (Selleck, S1623) and Apocynin (Selleck, S2425) were used as ROS inhibitors in this research.
For the singlet oxygen (1O2) detection, three samples of TC-WS-CQDs aqueous solution containing 1% 2,2,6,6-tetramethylpiperidine (TEMP) and one sample of 1% TEMP aqueous solution were treated by avoiding light for 30 min, 470 nm laser irradiation for 5 min, and 30 min respectively, and then transferred to quartz capillary for electron spin resonance (ESR) spectra measurement.
For the singlet oxygen (1O2) detection, three samples of TC-WS-CQDs aqueous solution containing 1% 2,2,6,6-tetramethylpiperidine (TEMP) and one sample of 1% TEMP aqueous solution were treated by avoiding light for 30 min, 470 nm laser irradiation for 5 min, and 30 min respectively, and then transferred to quartz capillary for electron spin resonance (ESR) spectra measurement.
10
Evaluating Rapamycin and Torin1 Effects
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Rapamycin and Torin1 were purchased from Beyotime Institute of Biotechnology (China). Chloroquine (CQ) and N-acetylcysteine (NAC) were purchased from Selleck Chemicals (USA). Hydroxyurea (HU) was from Sigma-Aldrich (USA). X-ray apparatus (X-RAD 225 OptiMAX) was from Precision X-ray (PXi).
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