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.

Abstract:

: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.

journal_name

J Cell Physiol

authors

Giese G,Kubbies M,Traub P

doi

10.1002/jcp.1041610204

subject

Has Abstract

pub_date

1994-11-01 00:00:00

pages

209-16

issue

2

eissn

0021-9541

issn

1097-4652

journal_volume

161

pub_type

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