Figure 1
From: Purifying Cytokinetic Cells from an Asynchronous Population
Purifying cytokinetic cells – strategy and practice.
(A) An illustration of mAG-Geminin dynamics in proliferating cells. mAG-Geminin signal drops at the metaphase-to-anaphase transition and starts accumulating again at the G1-S transition of the following cycle. During cytokinesis, mAG-Geminin levels are low. (B) mAG-Geminin dynamics in live L1210 cells. Time-lapse microscopy of a representative L1210 cell stably expressing mAG-Geminin. Images were taken at 3- min intervals using a ×20 air lens. mAG signal was normalized to maximum intensity at t = 0 (% of max). (C) (Top, left) mAG-Geminin-expressing L1210 cells were stained with Hoechst 33342 and their DNA distribution was measured by FACSAria III. (Top, right) A bivariate plot of mAG-Geminin vs. DNA. A distinct population of cells with 4 N DNA and low mAG-Geminin levels was gated (red circle). G2-early M cells and G1 cells were gated as well (gray and blue circles, respectively). (Bottom) DNA distributions of cytokinetic cells (red), G2-early M cells (gray) and the entire population (black) are shown.
