To those of you on the cell cycle board, I apologize for the redundancy, but I wanted some more input. We have been analyzing the cell cycle of 2 cell lines, a parent and a daughter stably transfected with a gene of interest. The transfectant has a significantly reduced doubling time compared to vector control. The most striking difference in DNA histograms is a fairly dramatic reduction in the percentage of G2/M cells in the transfectant (this is confirmed by integration with WinCycle - 0 order S phase and aggregate correction). There is a concomitant increase in G1 and S phase percentages. We feel that this indicates a decreased transit time across G2/M. Is this a correct assesment? We tried BrdU labeling to get an accurate picture, but both cell lines have very long doubling times (even the transfectant) and we were unable to get enough BrdU incorporated to visualize (We tried several times). Does our interpretation sound logical? Is there anything else we can or should do? We tried synchronization with aphidicolin but the cells did not progress through s and G2/M tightly enough to give conclusive data. Any comments or suggestions would be appreciated along with alternative hypothesis. Thanks John Tonkinson Lilly Research Labs JTonk@lilly.com
This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:43 EST