Hello,
I have carried out cell cycle assay of HeLa cells using Nocodazole(40ng/ml) and performed FACS of the samples collected at different time points. Since I am new to this technique, I feel a bit confused regarding the interpretation of the data. As far as I kbow, Nocodazole induce cell cycle arrest at G2/M boundary and enters uniformly to G1after release. I collected the cells at 0 time point to measure the success of arrest. I have attached a copy of the result at 0 time.What could be the reason why I got majority of the cells in G1 phase instead of G2/M. Is it due to the failure of nocodazole cell cycle arrest or does the way I am interpreting the results is wrong?.
I extremely appreciate your help and advice.
Thank you
Nocodazole is a microtubule poison that usually induces cell cycle arrest in prometaphase. However, it is also a down-regulator of Wnt-beta cathenin. When this activity predominates it may well be possible that cells do not enter in mitosis.
Thank you very much@Tomas Koltai.In that case still I should get cells in G2 phase though cells do not enter in to mitosis. Sorry if I am wrong about this.
I have the unproven idea that according to the concentration of nocodazole different effects may be achieved, including Wnt/Beta catenin downregulation. In order to get the CDK1 inhibitory effect I recommend you use the concentrations published in 2006. I am sending you the article.
How did your Asynchronous control population look? without looking at the control population it is difficult to assess the peak of Nocodazole arrested cells.
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