My substance decreases percentage of cell numbers in the G1 and increases percentage of cell numbers in the G2 phase while percentage of cell numbers in the S phase(cell cycle) remains same compared to the saline control in the cytometry assay. WB shows unregulated expression of Cycline D1.What can this suggest? (Btw, CCK-8 assay show higher proliferation rate in treated cells)
Hi Muhammad, we can really write a thesis here since with such little information the possibilities are endless.
I assume that the G1 phase is shorter and the G2 is longer. Or could it be G2/M arrest because there is more DNA damage.
- Can you tell something more about your substance? What is it?
- What is the cytometry assay? Propidium iodide staining so you are measuring the DNA content? Or are you using any other marker?
- What does unregulated expression mean - higher, lower, something else? Should we assume it's less in the G1 in your treated cells?
- Any chance you measured CDK4/6, pRb, cyclinE/cdk2 levels too?
- CCT8 assay is dependent on the level of dehydrogenases expressed by your cells. Could it be that your cells express more of these so you are getting a false positive increase in proliferation rate?
- If the cells really proliferate faster, do they survive later or do they accumulate damage and induce apoptosis?
Hello Muhammad, What cell numbers are you recovering from the different treatment groups? And how are you evaluating cell cycle eg DAPI, Ho, PI ? What about % apoptototic population in the different treatments is that changing between groups?
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