Figure 4

Promoter activity of the upstream sequence of functional eIF3f in quiescent A549 cells (G0), M, G1, and S synchronous cultures. A) Cell cultures in G0, M or S were transiently transfected with the promoter plasmid constructs and harvested 48 (for M and S cells) or 54 (for G0 cells) hours after transfection for luciferase activity assays. For cells in G1, cell cultures were transiently transfected with the promoter plasmid constructs for 20 hours, then arrested in M phase with nocodazole for 24 hours, and finally released from nocodazole to harvest cells 4 hours later for luciferase activity assays in G1. Luciferase activity with the empty vector pGL3 was below 10 RLU/μg of protein. Data expressed as mean ± SE; * p < 0.01 with respect to control (pGL3), n ≥ 3. B) Cell cycle distribution for nocodazole, HU and serum deprivation after release from arrest in M, S and G0, respectively. Cells were stained with DAPI at different timepoints to assess mitotic index. Micrographs of mitotic and interphase A549 cells are shown (top) at a magnification of 264×.