BEZ235, CAL-101, GSK650394, Roscovitine, Salubrinal, AZD8055, MK-2206 and PF4708671 were from Selleck Chemicals LLC (Houston, TX, USA). CN585 (6-(3,4-Dichlorophenyl)-4-(N,N-dimethylaminoethylthio)-2-phenyl-pyrimidine) was from Merck Millipore (Merck, Damstadt, Germany). Ionomycin, PMA, CsA, FK-506 and BV-02 were from Sigma-Aldrich.
Salubrinal
Manufactured by Selleck Chemicals
Sourced in United States, China
Salubrinal is a laboratory chemical compound used in research. It functions as a selective inhibitor of the protein phosphatase 1 (PP1) enzyme, which plays a role in the regulation of cellular stress responses.
Automatically generated - may contain errors
Lab products found in correlation
Isrib, by Selleck Chemicals (2 mentions)
Eif2α, by Cell Signaling Technology (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Fk506, by Merck Group (1 mentions)
Cal 101, by Selleck Chemicals (1 mentions)
Gsk650394, by Selleck Chemicals (1 mentions)
Roscovitine, by Selleck Chemicals (1 mentions)
Pf 4708671, by Selleck Chemicals (1 mentions)
Bez235, by Selleck Chemicals (1 mentions)
Azd8055, by Selleck Chemicals (1 mentions)
Mk 2206, by Selleck Chemicals (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Thapsigargin, by Santa Cruz Biotechnology (1 mentions)
Glutamine, by Merck Group (1 mentions)
Jnk inhibitor sp600125, by Selleck Chemicals (1 mentions)
Hepes, by Merck Group (1 mentions)
Du145, by American Type Culture Collection (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
11 protocols using salubrinal
1
Chemical Inhibitors of Cell Signaling Pathways
2
Cell Line Culturing and Inhibitor Treatments
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The prostate cancer cell lines PC-3 and DU-145 were purchased from the ATCC; Dr Vivek Rangnekar, University of Kentucky, Lexington gifted Par-4+/+ and Par-4−/− MEFs; Dr Bert Vogelstein, Johns Hopkins University, Baltimore, MD gifted p53+/+ HCT-116 and p53−/− HCT-116 cell lines. These cell lines were cultured in RPMI 1640/DMEM medium (Invitrogen Life Technologies, Carlsberg, CA, USA) supplemented with 10% fetal bovine serum, 70 mg/l penicillin, 100 mg/l streptomycin, 6 mm HEPES and 2 mm l -glutamine (Sigma-Aldrich, St Louis, MO, USA) at 37 °C and 5% CO2. The major inhibitors including ER stress inhibitor Salubrinal, JNK inhibitor SP600125 were purchased from Selleck Chemicals, Houston, TX, USA. AKT inhibitor AT7867, caspase inhibitor Fmk-z-VAD and ER stress inducer Thapsigargin were procured from Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA.
3
Metabolic Regulation via GDF15 Signaling
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2-DG (#S4701), ISRIB (#S0706), Salubrinal (#S2923), GCN2IB (#S8929), C16(#S9668), GSK2656157 (#S7033), and Compound C (#S7840) were purchased from Selleck (shanghai, China). Lipopolysaccharide (LPS, #L2630, E. coli 0111:B4) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Mouse recombinant GDF15 (#10596-GD-025) was purchased from R&D Systems (Minnesota, USA). Pooled mouse ATF4 siRNA (#1. Sense-5’-CUCCCAGAAAGUUUAAUAATT-3’, anti-sense-5’- UUAUUAAACUUUCUGGGAGTT-3’; #2. Sense-5’-GCUGCUUACAUUACUCUAATT-3’, anti-sense-5’-UUAGAGUAAUGUAAGCAGCTT-3’; #. Sense-5’-GUCUCUUAGAUGACUAUCUTT-3’, anti-sense-5’-AGAUAGUCAUCUAAGAGACTT-3’) and pooled mouse GDF15 siRNA (#1. Sense-5’-CUCGAACUCAGAACCAAGUTT-3’, anti-sense-5’-ACUUGGUUCUGAGUUCGAGTT-3’; #2. Sense-5’-GUGGUUCUUAUGCACAGGATT-3’, anti-sense-5’-UCCUGUGCAUAAGAACCACTT-3’; #3. Sense-5’-CUGCUAAUAAAGGUGAGCUTT-3’, anti-sense-5’-AGCUCACCUUUAUUAGCAGTT-3’) were synthesized by GenePharma (Shanghai, China). Antibodies against phosphorylated eIF2α (3597 S), total eIF2α (5324 S), ATF4 (11,815 S), phosphorylated AMPK (2535 S), and AMPK (2532 S) were purchased from cell signaling technology (MA, USA). Antibodies against Glut1(66,290), HK2 (66,974), PFKFB3 (13,123), and PKM2 (60,268) were obtained from Proteintech (Wuhan, China) Antibody against GDF15 (ab105738) was purchased from Abcam (MA, USA).
4
Palmitic Acid-Induced Stress Signaling
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All chemicals and buffers were of analytical grade and purchased from ThermoFisher Scientific (Massachusetts, USA). Palmitic acid (A3803) and fatty acid free bovine serum albumin (BSA, P5585) were obtained from MilliporeSigma, St. Louis, MS. 15-deoxy-Δ12,14-Prostaglandin J2 (PGJ2) and GSK2606414 (GSK260) were purchased from Cayman, Ann Arbor, MI. SP600125 (JNKi), STF-083010 (IRE1αi, endonuclease inhibitor), Salubrinal (eIF2αi, dephosphorylation inhibitor), U0126 (ERKi, MEK1/2 or ERK1/2 Inhibitor) were obtained from Selleck Chemicals, Houston, TX.
5
Synthesis and Antibody Analysis of IMB-6G
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IMB-6G was synthesized according to previously described methods [18 (link)]. Antibodies against Bip, CHOP, PERK, IRE1α, eIF2α, phospho-PERK (Thr980), phospho-eIF2α (Ser51), phospho-ASK1 (Thr845), phospho-JNK (Thr183/Tyr185), Bax and Bad were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-phospho-IRE1α (Ser724) and ATF6 antibodies were from Abcam (Cambridge, MA, USA). Anti-Bim, PUMA and Cytochrome c antibodies were purchased from Santa Cruz (Santa Cruz, CA, USA). SP600125, NQDI-1, Thapsigargin, MTT and β-actin antibody were obtained from Sigma (St. Louis, MO, USA). Salubrinal (ER stress inhibitor) was purchased from Selleckchem (Shanghai, China).
6
Antibody and Small Molecule Profiling for ER Stress Response
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Antibodies including HT-7 (catalog no.: MN1000; Invitrogen), AT8 (catalog no.: MN1020; Invitrogen), YFP (catalog no.: ab6556; Abcam), HSP90 (catalog no.: ab13492; Abcam), lamin A/C (catalog no.: 2032; Cell Signaling Technology), GAPDH (catalog no.: ab8245; Abcam), T-PERK (catalog no.: 3192; Cell Signaling Technology), phosphorylated PERK (catalog no.: 3179; Cell Signaling Technology), eIF2α (catalog no.: 5324; Cell Signaling Technology), p-eIF2α (catalog no.: 5324; Cell Signaling Technology) were pretested to detect the targeted proteins. Small molecules included GSK2656157 (catalog no.: 9466-5; BioVision), GSK2606414 (catalog no.: S7307; Selleckchem), salubrinal (catalog no.: S2923; Selleckchem), ISRIB (catalog no.: S7400; Selleckchem); Ceapin-A7 (catalog no.: SML2330; Sigma–Aldrich), 4u8C (catalog no.: CAS14003-96-4; Calbiochem), and AA147 (product no.: 6538059; ChemBridge) were prepared in dimethyl sulfoxide following the manufacturer’s instruction and stored at −80 °C as stock solution. ER stress–inducing chemical, thapsigargin, was dissolved in dimethyl sulfoxide and added to the cell culture media at a concentration of 300 nM. The working solutions were freshly prepared with −80 °C stock solution.
7
Hepatic Stellate Cell Fibrosis Model
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The human hepatic stellate cell line LX2 was cultured in modified Eagle’s medium (MEM) supplemented with 10% (v/v) fetal bovine serum (FBS) (Gibco, Carlsbad, CA, United States). The cells were kept at 37°C in a humidified incubator with 5% CO2. PDGF-BB (Sigma no. P4056) was purchased from Sigma-Aldrich (St. Louis, MO, United States). Tunicamycin was purchased from Dalian Meilun Biological Technology Co., Ltd. (Dalian, China) and salubrinal was purchased from Selleck (Houston, TX, United States). The in vitro model of fibrosis was established by treating LX2 cells with PDGF-BB, and then LX2 cells were incubated in the absence or presence of SalA (25 μM) for 24 h, which were determined by preliminary experiment and reported (Lin et al., 2006 (link); Ding et al., 2016 (link)). In vitro, LX2 cells were stimulated with 20 ng of PDGF-BB for 24 h. After incubation with 10 μM of EX527 for 6 h, the LX2 cells were subjected to 20 ng of PDGF-BB and/or 25 μM of SalA stimulation as needed.
8
Temporal Regulation of ER Stress-Induced Apoptosis in TBI
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The first experiment analyzed ROS production and expression of ER stress- and apoptosis-related proteins in the injured cortex at different time points. Rats were divided into six subgroups including a sham control group and groups sacrificed at 1, 3, 6, 12, and 24 hours post-TBI (n = 6/group). The second experiment was performed to study the effect of the ER stress inhibitor, Salubrinal (Sal), post-TBI. Rats were randomized into four groups including sham control, TBI, sham + Sal, and TBI + Sal groups. Salubrinal (1 mg/kg) (Selleck, Houston, TX, USA) was injected intraperitoneally 30 minutes before TBI (Nakka et al., 2010; Zhang et al., 2015) (n = 6/group).
9
Pathogen-free Mouse Model Study
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Eight-week-old male DBA/1J mice and six-week-old male C57BL/6 mice (Shanghai SLAC Laboratory Animal Co., Ltd., Shanghai, China) were maintained under pathogen-free conditions at Shanghai Jiao Tong University School of Medicine. All experimental procedures were performed in accordance with the guidelines of the Animal Care and Use Committee. Salubrinal and MG132 were purchased from Selleck Chemicals (Houston, TX, USA). BAY11 was purchased from Beyotime Biotechnology (Shanghai, China), and cycloheximide (CHX) was purchased from MedChemExpress (Monmouth Junction, NJ, USA). Chloroquine (CQ) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
10
Salubrinal Administration for Treatment
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Salubrinal (Selleck) was reconstituted in phosphate buffered saline (PBS) containing 1% DMSO, and the solution was filtered using a 0.2 μm sterile syringe filter. Then, 0.5 mg/kg·Salubrinal was administered by intraperitoneal injection once every day for 4 weeks. The dose was determined based on previous research (Rani et al., 2017 (link)).
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