Comprehensive and quantitative analysis of G1 cyclins. A tool for studying the cell cycle
Fig 3
Cyclin waves in S. cerevisiae growing in normal lab conditions (YPD at 30°C).
G1 cyclin waves as determined in this research. From top to bottom, the panels show the Cdc28 G1 cyclins, the Pho85 G1 cyclins and the different molecular markers (Clb5 and Sic1) and morphological markers (budding percentage and size for elutriation). START, determined as the moment in which Sic1 and Clb5 amounts were identical, is extrapolated as a dashed line to all the panels. A) Cells were synchronized by centrifugal elutriation (left panels) or α-factor (right panels). Time 0 corresponds to the moment of cell removal from the elutriation device or α-factor removal, after which cells were incubated under agitation at 30°C. Note that the time elapsed before the cells resumed the cell cycle was greater after elutriation than after α-factor treatment. B) Since the amount of Cln3-3HA is very low, it is plotted both with the other Clns and individually so as to clearly represent levels. In the case of α-factor synchronization, at least three independent western blot experiments as in Fig 2B were quantified, standardized using loading control and relativized to the maximum expression amounts (Cln2). Values are expresed as mean±SEM.