Skip to main content
. Author manuscript; available in PMC: 2013 Jul 13.
Published in final edited form as: Mol Cell. 2012 May 31;47(1):50–60. doi: 10.1016/j.molcel.2012.04.028

Figure 3. Flow cytometric quantitation of S-phase progression after MNNG and BzNU treatment.

Figure 3

Cdc10-M17 cells grown at 25°C and synchronized in G1 by shifting to 37°C for 4.5 hours. Before release from arrest, cells were treated with BNNG or MNNG (10 min), centrifuged and resuspended in YEL. Samples were taken every 20 min and nuclear DNA content was measured by flow cytometry.

(A) Overlays of S-phase flow cytometry histogram stacks and

(B) S-phase progression after treatment with high concentrations of MNNG leading to ~0.5% cell survival. S-phase progression (%) was measured by the relative shifting of the mean of S-phase peaks from unreplicated 1C toward fully replicated 2C values preset as 0% and 100% respectively.

(C) Overlays of S-phase flow cytometry histogram stacks and

(D) S-phase progression comparing WT (SP413), atl1 (GM62), rhp26 (GM106), rhp26 atl1 (GM107), swi10 (GM108), swi10 atl1 (GM109) strains after treatment with BzNU (0.5 mg/mL). Values are the mean of triplicate experiments and error bars represent SEM. The values of cell survival were as follows: wild-type (27%), atl1 (32.7%), rhp26 (13%), rhp26 atl1 (17.5%), swi10 (8.8%), swi10 atl1 (25.8%). See also Figure S3.