I tested a natural product to treat breast cancer cell line. The cells was treated 24h and I used PI to dye the cells. I also used Doxorubicin as the control group. From the look, I can see that the natural product did not have much effect on cancer cell nor arrest the progression but the SubG1 population was slightly increased. I get the overview idea of cell cycle test but I'm not sure I read it right. (I'm new in this topic). If anyone has done this please let me know if I'm in the right direction. I attached the result of cell cycle as well. Thanks!!
Dear Aisyah,
Your data is fine and I can see a clear difference between the histograms of the Untreated cells and your natural product treated cells (S-phase increase) apart from the slight increase in the SubG1. The only problem here is in the analysis. If you use the same marker regions for your test sample that you have used for the Untreated you will get an increase in the S-phase population and a subsequent decrease in the other regions.
I don't think you need to change the marker regions for each test sample which leads to misinterpretation of flow data. The reason you run controls is to help you in proper gating.
I can re-analyze it if you send me the FCS files.
The markers should be fixed according to the control and applied to the rest. I do not have much experience in flowcytometry but I did carried out some and it seems like the thresholds for A, B, C and D differs because they are skewed to the left. Did you set up the setting/ calibrate with control first (about 200 and 400) and run the treated at the same time/day after that? But galangal does seem to increase S population although you only showed one time point which might not really show the effect. And I have a question about staining as I had also done only PI staining but is the sub G1 real without doing PI-annexin V staining. I am also still learning about this
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