HMN-154 interacts with NF-YB and thereby interrupts the binding of the NF-Y heterotrimer to DNA. NF-YB and thymosin β-10 are specific cellular binding proteins of HMN-154 and that this shared region is necessary for the binding to HMN-154. HMN-154 inhibits DNA binding of NF-Y to the human major histocompatibility complex class II human leukocyte antigen DRA Y-box sequence in a dose-dependent manner. HMN-154 shows very strong cytotoxicity against KB and colon38 cells with an IC50 value of 0.0026 and 0.003 μg/mL, respectively. HMN-154/BSA binds recombinant NF-YB or thymosin β-10 and the binding is inhibited by the addition of HMN-154 as the competitor. The binding between HMN-154 and NF-YB is specific and depends on its cytotoxicity[1].
Kinase Assay
Cell Assay
Cells are seeded into a 96-well microplate at a cell density of 10000/well. Drug is added on the next day, and the plate then is incubated for 72 h at 37°C. The growth inhibitory concentration is measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay[1].
Animal Administration
References
[1]. Tanaka H, et al. Isolation of cDNAs encoding cellular drug-binding proteins using a novel expression cloning procedure: drug-western. Mol Pharmacol. 1999 Feb;55(2):356-63.
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