关键词: Sf9细胞系； 组蛋白H3； 磷酸化, 有丝分裂, 免疫荧光标记, 染色质凝缩

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.

Key words: Sf9 cell line, histone H3, phosphorylation, mitosis, immunofluorescence labeling, chromosome condensation