High resolution analysis of cell cycle-correlated vimentin expression in asynchronously grown, TPA-treated MPC-11 cells by the novel flow cytometric multiparameter BrdU-Hoechst/PI and immunolabeling technique.
High resolution, multiparameter analysis using the flow cytometric BrdU/Hoechst quenching technique has been applied to study cell cycle kinetics and vimentin expression in individual cells of asynchronously grown MPC-11 mouse plasmacytoma cell cultures treated with 12-O-tetradecanoylphorbol-13-acetate (TPA) to induce in vitro differentiation. BrdU treatment up to 16 h in the absence or presence of TPA did not affect either cell cycle progression or the kinetics or quantity of vimentin expression. TPA-treated cells became arrested in G1 phase of the second cell cycle; however, this G1 phase arrest was transient only. In addition, G1 phase cells located prior to a putative transition point at the beginning of TPA treatment were completely blocked in cell cycle progression. There is also evidence that cells located in G1 or G2/M phase at the beginning of TPA treatment finally expressed low levels of vimentin. On the contrary, cells located in S phase at TPA exposure showed high vimentin levels after treatment. The results presented here show that, with the flow cytometric BrdU/Hoechst quenching technique, one can correlate time-dependent protein expression at the single cell level in asynchronously grown cultures not only with the actual cell cycle state, but also with the history of cell replication.
Giese
G
G
Max-Planck-Institut für Zellbiologie, Ladenburg, Germany.
Kubbies
M
M
Traub
P
P
D000818:Animals; D006690:Bisbenzimidazole; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002453:Cell Cycle; D002455:Cell Division / Q000187:drug effects; D005434:Flow Cytometry / Q000379:methods*; D007158:Immunologic Techniques; D007834:Lasers; D051379:Mice; D010954:Plasmacytoma / Q000378:metabolism* / Q000473:pathology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000378:metabolism*
D014746:Vimentin; D001973:Bromodeoxyuridine; D006690:Bisbenzimidazole; D013755:Tetradecanoylphorbol Acetate