Effect Of The Subcellular Location Of DAXX On Apoptosis Of Macrophage Cells Induced By Ox-LDL

Objective: Aimed to evaluate the effects of subcellular localization of DAXX onox-LDL induced the apoptosis of RAW264.7cells.Background: The fuction of DAXX is different because of its different location,though the singnaling of apoptosis regulated by DAXX is well known. The role ofDAXX, promote or hinder apoptosis, its still remains controversial.Methods: RAW264.7cells were transient transfected with DAXX (WT), DAXX(S667A) mutant, DAXX(W621A) mutant or control vector respectively. These cellswere cultured with normal conditions for24h, and then incubated with100μg/mLox-LDL for48h.Immunofluorescent assay was used to assay the localization of DAXX.MTT and FCM was used to determine cellular viability and apoptosis. RT-PCR andWestern Blotting were used to analyze the expression of mRNA and proteinrespectively.Results: The Results of Immunofluorescent assay demonstrated that the DAXX(S667A) is particularly localized to nucleus and the DAXX (W621A) localized tocytoplasm. Overexpression of DAXX (W621A) promoted cell apoptosis induced byox-LDL via activation of ASK-1and JNK, which same as DAXX (WT) groups.Conversely, Overexpression of DAXX (S667A) protected cells.Conclusion: The subcellular localization of DAXX determines its role in RAW264.7cellular apoptosis, which maybe related to the ASK-JNK singling pathway.