http://en.dogeno.us/2008/04/synchronize-cell-cycle-in-culture/ thoughts about life and science, blogged by Liang Cai | cail.cn Wed, 08 Feb 2012 04:54:25 +0000 hourly 1 http://wordpress.org/?v=3.3.1 http://en.dogeno.us/2008/04/synchronize-cell-cycle-in-culture/#comment-17565 cail Sun, 11 Oct 2009 02:19:04 +0000 http://en.dogeno.us/?p=1588#comment-17565 this post is the top view in my "experiment" category and the number two in google search this post is the top view in my "experiment" category and the number two in google search

]]> http://en.dogeno.us/2008/04/synchronize-cell-cycle-in-culture/#comment-15209 cail Mon, 05 May 2008 20:24:29 +0000 http://en.dogeno.us/?p=1588#comment-15209 A recent paper "MOBKL1A/MOBKL1B Phosphorylation by MST1 and MST2 Inhibits Cell Proliferation" >> in result Nocodazole arrests cells at metaphase by disruption of the spindle structure and, thus, activates the spindle checkpoint; to determine whether MST1/MST2 activation occurs in the course of a normal mitosis, we collected cells easily shaken off plates of freely cycling cultures. The presence of robust histone H3 (Ser10) phosphorylation in these cells, as compared to freely cycling, adherent cells, provided an indicator that a substantial fraction of the cells collected by shake-off were in mitosis (Figure 4D, third row from top). >> in method U2OS cells were plated one day before the onset of synchronization, which was achieved by growing the cells in their complete medium containing 0.4 mg/ml Nocodazole (Sigma) for 16 hr. The rounded U2OS cells then were either collected (shaken off) and used as mitotic samples or washed twice with fresh medium and replated for different time points needed for studying the mitotic exit. A recent paper "MOBKL1A/MOBKL1B Phosphorylation by MST1 and MST2 Inhibits Cell Proliferation"
>> in result
Nocodazole arrests cells at metaphase by disruption of the spindle structure and, thus, activates the spindle checkpoint; to determine whether MST1/MST2 activation occurs in the course of a normal mitosis, we collected cells easily shaken off plates of freely cycling cultures. The presence of robust histone H3 (Ser10) phosphorylation in these cells, as compared to freely cycling, adherent cells, provided an indicator that a substantial fraction of the cells collected by shake-off were in mitosis (Figure 4D, third row from top).
>> in method
U2OS cells were plated one day before the onset of synchronization, which was achieved by growing the cells in their complete medium containing 0.4 mg/ml Nocodazole (Sigma) for 16 hr. The rounded U2OS cells then were either collected (shaken off) and used as mitotic samples or washed twice with fresh medium and replated for different time points needed for studying the mitotic exit.

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