선택한 단어 수는 입니다.
최소 단어 이상 선택하여야 합니다.
최소 단어 이상 선택하여야 합니다.
선택한 단어 수는 30입니다.
최대 10 단어까지만 선택 가능합니다.
최대 10 단어까지만 선택 가능합니다.
제안 방법
- In subsequent experiments, kinetic analyses based on the Michaelis-Menten equation were performed with ethyl-3,4- dephostatin and DUSP22 to provide experimental evidences for the mechanism of DUSP22 catalysis and for binding of the inhibitor to the active site of the phosphatase. The Km value of DUSP22 for OMFP was 31.
- Next, the in vivo effect of ethyl-3,4-dephostatin on DUSP22 activity were determined by measuring the phosphorylation status of endogenous JNK. HEK 293 cells were transfected with FLAG-DUSP22 expression plasmid, pretreated with various concentrations of ethyl-3,4-dephostatin for 3 h, and then the levels of phospho-JNK were determined with Western blotting analysis.
대상 데이터
- Anti-phospho-JNK (specific for phospho-Thr183 and phospho-Tyr185), anti-p38, antiphospho-p38 (Thr-180/Tyr-182), anti-ERK1/2, and antiphospho-ERK1/2 (Thr-202/Tyr-204) antibodies were purchased from Cell Signaling Technology (Danvers, MA). AntiJNK antibody was from Santa Cruz Biotechnology (Santa Cruz, CA). Ethyl-3,4-dephostatin, 3-Omethylfluorescein phosphate (OMFP), anti-FLAG M2 antibody, and antiFLAG M2 agarose beads were from Sigma-Aldrich (St.
- FLAG-tagged DUSP22 wild-type and DUSP22 DACS mutant were constructed in pcDNA3.1 (Invitrogen). His-tagged DUSP22 WT was constructed in pET28a plasmid (Novagen, Darmstadt, Germany) for protein expression in Escherichia coli.
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