Dynamic distribution of histone H3 Ser10 phosphorylation in <em>Spodoptera frugiperda</em> Sf9 cells during mitosis
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ACTA ENTOMOLOGICA SINICA  2017, Vol. 60 Issue (11): 1278-1284    DOI: 10.16380/j.kcxb.2017.11.005
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Dynamic distribution of histone H3 Ser10 phosphorylation in Spodoptera frugiperda Sf9 cells during mitosis
WANG Jiao-Ling, MA Ya-Fei, WANG Shuang, KAN Yun-Chao, ZHANG Bing*
(Henan Key Laboratory of Insect Biology in Funiu Mountain, Henan Provincial Engineering Laboratory of Insects Bio-reactor, China-UK-NYNU-Res Joint Laboratory of Insect Biology, Nanyang Normal University, Nanyang, Henan 473061, China)
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Abstract 【Aim】 To explore the function of histone H3 Ser10 phosphorylation, an epigenetic marker, in cells of Spodoptera frugiperda cell line Sf9 during mitosis. 【Methods】 The sequence conservation of histone H3 was analyzed by sequence alignment. Antibody was prepared by synthesis of histone H3 pepetide RK(pS)TGGKAPRKQLC, in which the tenth serine is specifically phosphorylated by solid phase method. Sf9 cells were cultured, and the mitotic cells were prepared by cell climbing. The numbers of cells in different mitotic stages were counted, and then the localization characteristics of histone H3 Ser10 phosphorylation antibody in these stages were detected by cytology immunofluorescence. 【Results】 Sequence alignment analysis revealed that histone H3 1st-60th amino acids were highly conserved in most species. In Sf9 cells, histone H3 Ser10 phosphorylation began to appear near the nuclear membrane with a punctate distribution pattern in early prophase, and spread throughout the whole chromosome in early metaphase during the cell cycle. During anaphase, the fluorescent signals of Ser10-phosphorylated H3 disappeared from chromosomes, and the dephosphorylation completed in telophase. 【Conclusion】 Histone H3 Ser10 phosphorylation is associated with chromosome condensation in Sf9 cells during mitosis.
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WANG Jiao-Ling
MA Ya-Fei
WANG Shuang
HAN Yun-Chao
Key wordsSf9 cell line   histone H3   phosphorylation   mitosis   immunofluorescence labeling   chromosome condensation     
Cite this article:   
WANG Jiao-Ling,MA Ya-Fei,WANG Shuang et al. Dynamic distribution of histone H3 Ser10 phosphorylation in Spodoptera frugiperda Sf9 cells during mitosis[J]. ACTA ENTOMOLOGICA SINICA, 2017, 60(11): 1278-1284.
http://www.insect.org.cn/EN/10.16380/j.kcxb.2017.11.005      or     http://www.insect.org.cn/EN/Y2017/V60/I11/1278
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