Fig 8.

N40 expression has little to no effect on cell cycle progression and DNA synthesis. (A) Cell cycle analysis of C33A cells transiently expressing YFP alone or YFP-tagged WT or WF N40. The DNA content was determined by Hoechst staining and flow cytometry. For each transfection, the cells were either grown asynchronously (Asynch.) or synchronized with mimosine for 24 h (Mimo. block) and then released in nocodazole for another 24 h (Released in nocodazole). (B) BrdU incorporation of C33A cells transiently expressing YFP alone or YFP-tagged WT or WF N40. Cells were pulsed for 1 h with BrdU 48 h posttransfection and were then dually labeled for BrdU and DNA using an APC-coupled antibody and 7-AAD staining, respectively. For each sample, BrdU incorporation is graphed as a function of the DNA content. BrdU incorporation of cells treated with aphidicolin is shown as a negative control. Quantification of the percentage of cells incorporating BrdU, from three independent experiments, is shown on the right (average ± standard deviation).