MDS and AML cell lines were cultured in the presence of DMSO, LEN (S1029; Selleckchem), or POM (P0018; Sigma) at the indicated concentrations for the apoptosis studies and the intracellular calcium analysis. MDSL cells were treated with DMSO or 10 μM LEN for 2 d, followed by cotreatment with 20 μM Lsx (Neuronal Differentiation Inducer III; 480743; EMD Millipore) for 1 d, 400 nM ionomycin (Calcium Ionophore, C5722, Sigma) for 1 d, 200 μM EGTA (045172; Fisher) for 5 d, or 5 μM BAPTA-AM (A1076; Sigma) for 5 d for the apoptosis assay. MDSL and TF-1 cells were treated with DMSO or 10 μM LEN for 2 d, followed by cotreatment with 5 μM N-phenylmaleimide (P27100; Sigma), 20 μM Z-VAD-FMK (a pan-caspase inhibitor; FMK001; R&D Systems), 1 μM E-64 (E3132; Sigma), 1 μM pepstatin (P5318; Sigma), and 100 μM PD150606 (D5946; Sigma) for 3 d for the apoptosis assay. MDSL cells were treated with 1 μM POM for 2 d, followed by cotreatment with 300 μM PD150606 for 3 d for the apoptosis assay. MDS and AML cells were treated with DMSO or 10 μM LEN for 4 d, or with 5 μM ionomycin for 30 min, for the measurement of calpain activity.
S1029
Manufactured by Selleck Chemicals
S1029 is a laboratory equipment product. It serves a core function in scientific research and analysis, but a detailed description cannot be provided in an unbiased and factual manner without potential extrapolation or interpretation.
Automatically generated - may contain errors
Lab products found in correlation
Calcium ionophore, by Merck Group (1 mentions)
Pepstatin, by Merck Group (1 mentions)
P5318, by Merck Group (1 mentions)
P0018, by Merck Group (1 mentions)
Pd150606, by Merck Group (1 mentions)
Fmk001, by R&D Systems (1 mentions)
Ethylene glycol tetraacetic acid (egta), by Thermo Fisher Scientific (1 mentions)
Bapta am, by Merck Group (1 mentions)
S7067, by Merck Group (1 mentions)
2 protocols using s1029
1
Apoptosis and Calcium Signaling Assays
2
Inhibition of p38 MAPK and TNF-α Secretion
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Specific inhibitor of p38 MAPK, SB202190 (10 μM, S7067, Sigma) and SB203580 (10 μM, T1764, TargetMol), and a highly selective TNF-α secretion inhibitor lenalidomide (CC-5013) (10 μM, S1029, Selleckchem) were all dissolved in dimethyl sulfoxide (DMSO), subsequently diluted in 0.1 M phosphate-buffered physiological saline (PBS) and sterilized through a 0.22-µm filter. For in vivo study, SB202190 (25 μg/kg/d), SB203580 (50 mg/kg/d) or lenalidomide (50 mg/kg/d) was pretreated intraperitoneally once a day on 3 consecutive days and 2 h ahead of HF-IR as well. SB202190 and SB203580 was injected respectively, and 0.1% DMSO was used as solvent control.
In addition, recombinant murine TNF-α/TNFSF2 (P6020, Beyotime, China) at a working concentration of 1.0 ng/ml was administrated to TM4 Sertoli cells in vitro for different times.
In addition, recombinant murine TNF-α/TNFSF2 (P6020, Beyotime, China) at a working concentration of 1.0 ng/ml was administrated to TM4 Sertoli cells in vitro for different times.
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