ABSTRACT
ANNA GRÉEN, BETTINA SARG, HENRIK GRÉEN, ANITA LÖNN, HERBERT H. LINDNER, and INGEMAR RUNDQUIST
6.1 Introduction .................................................................................. 157 6.2 Methods ....................................................................................... 159 6.2.1 Isolation of Peripheral Blood Lymphocytes ...................... 159 6.2.2 Activation of Peripheral Blood Lymphocytes,
Cell Culture, and Staining ............................................................ 159 6.2.3 Jurkat Cell Culture and Staining ........................................ 159 6.2.4 T Cell Assessments ............................................................ 160 6.2.5 The CFSE Staining and Flow Cytometry Measurements .. 161 6.2.6 CD3 Staining and Flow Cytometry Measurements ........... 161 6.2.7 Cell Cycle Analysis Using PI ............................................. 162 6.2.8 Detection of Apoptotic Peripheral Blood Lymphocytes .... 162 6.2.9 Cell Sorting ........................................................................ 162 6.2.10 Capillary Electrophoresis ................................................. 164 6.2.11 Reversed Phase High Performance Liquid
Chromatography .......................................................................... 164 6.3 Discussion .................................................................................... 164 6.4 Results .......................................................................................... 169 6.4.1 T Cell Activation Results in Rapidly Proliferating T Cell
Populations ................................................................................... 169
6.4.2 H1.5 Expression is Increased in Proliferating T Cells Compared with Resting Lymphocytes ......................................... 171
6.4.3 Proliferating T Cells and Jurkat Cells Contain Multiple Phosphorylated H1 Subtypes ....................................................... 173
6.4.4 Flow Sorting of T Cells and Jurkat Cells in Different Cell Cycle Phases ................................................................................ 175
6.4.5 The Phosphorylation of H1 Histones Starts in the G1 Phase of the Cell Cycle in Normal Proliferating T Cells ............ 181
6.4.6 Exponentially Growing Jurkat Cells Contain More Extensively Phosphorylated H1 Subtypes in the G1 Phase of the Cell Cycle Compared with Activated T Cells ................... 183
6.5 Conclusion ................................................................................... 186 Keywords .............................................................................................. 186 Acknowledgment .................................................................................. 186 Authors’ Contributions .......................................................................... 187 References ............................................................................................. 187 Credits ................................................................................................... 189
6.1 INTRODUCTION
Cell division is a complex process, in which correct passage through the cell cycle is essential for cell survival and correct transmission of genetic information to the daughter cells. During the cell cycle, the cell nucleus undergoes dramatic structural changes. DNA, which is compacted into chromatin by various proteins, is locally decondensed in S phase but condenses in prophase. In metaphase, highly condensed chromosomes are visible which start to segregate during anaphase. Segregation is completed during telophase, and two daughter cells are produced. Before re-entry into G1, the chromatin again becomes dispersed.
