ABSTRACT
The cell cycle is a fundamental process essential for cell proliferation, differentiation and development. It consists of four major phases: G1, S, G2 and M. These phases collectively drive the reproductive cycle and are meticulously regulated by various proteins that play crucial roles in both the prevention and progression of cancer. Traditional methods for studying these functions, such as flow cytometry, require a substantial number of cells to ensure accuracy. In this study, we have developed a user-friendly immunofluorescence-based method for identifying cell cycle stages, providing single-cell resolution and precise identification of G1, early/mid S, late S, early/mid G2, late G2, and each sub-stage of the M phase using fluorescence microscopy called ImmunoCellCycle-ID. This method provides high-precision cell cycle identification and can serve as an alternative to, or in combination with, traditional flow cytometry to dissect detailed sub-stages of the cell cycle in a variety of cell lines.
Footnotes
Author contributions
Conceptualization: A.S.; Data curation: Y-L.C.; Formal analysis: Y-L.C.; Funding acquisition: A.S.; Methodology: Y-C.C.; Supervision: A.S.; Validation: Y-L.C.; Writing – original draft: A.S.; Writing – review & editing: Y-L.C., Y-C.C.
Funding
This work is supported by the Wisconsin Partnership Program, Research Forward from the University of Wisconsin-Madison Office of the Vice Chancellor for Research with funding from the Wisconsin Alumni Research Foundation, start-up funding from University of Wisconsin-Madison SMPH, UW Carbone Cancer Center, and McArdle Laboratory for Cancer Research and National Institutes of Health (NIH; grant R35GM147525 and U54AI170660 to A.S.). Deposited in PMC for release after 12 months.
Data availability
All relevant data can be found within the article and its supplementary information. Other data and original images used in this study are available from the corresponding author upon reasonable request.
