Cyclin B1 expression in HeLa S3 cells studied by flow cytometry.
Using a procedure to stain cells simultaneously for cyclin B1 protein and DNA, we have examined cyclin B1 expression by flow cytometry in human cells under a variety of perturbing and nonperturbing conditions. The method described is useful for measuring relative differences in cyclin B level (immunochemically detectable epitope) as a function of cell cycle position on an individual cell basis and thus to examine cell cycle-related changes in cyclin B expression without prior cell synchronization. We show that in HeLaS3 cells, cyclin B1 accumulates in cells only after they become 4C and have resided in G2 for a short period of time. During colcemid-induced mitotic arrest cyclin B1 continues to accumulate in HeLa S3 cells, and under specific conditions of aphidicolin-induced unbalanced cell growth induced, cyclin B accumulates to supranormal levels prior to mitosis. Flow cytometric analysis of cyclin B expression and DNA content permits detailed examination of the effects of cell cycle perturbations on cyclin B expression under a variety of conditions.
Sherwood
S W
SW
Department of Biological Sciences, Stanford University, California 94305.
Rush
D F
DF
Kung
A L
AL
Schimke
R T
RT
D016590:Aphidicolin / Q000494:pharmacology; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D016213:Cyclins / Q000235:genetics* / Q000378:metabolism*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D003703:Demecolcine / Q000494:pharmacology; D005434:Flow Cytometry; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans
D016213:Cyclins; D016590:Aphidicolin; D004247:DNA; D003703:Demecolcine