Er ccd camera

TO-HCT116 and ORC2_H-2 cells were seeded in ibidi µ-Slide 8-Well Glass Bottom, in the presence or absence of 0.75 μg/mL doxycycline for 24 hr. Next, 2 mM thymidine was added and samples incubated ±dox for 24 hr. Two hours prior to washing out thymidine, 500 nM auxin were added to the dox treated wells. Samples were then imaged beginning 4 hr after thymidine release and the timepoints reconstructed from time-lapse images using volocity software. Images were acquired approximately every 5 min on a Perkin Elmer spinning disc confocal equipped with a Nikon-TiE inverted microscope using 40X objective lens with an Orca ER CCD camera. Images presented are maximum intensity projections of a z-stack (z = 3 μM).

Nuclei were fixed and stained with DRAQ5 Fluorescent Probe Solution as per the manufacturer’s guidelines (ThermoFisher #62251). Images were taken using a Perkin Elmer spinning disc confocal equipped with a Nikon-TiE inverted microscope using 60X objective lens with an Orca ER CCD camera. Images presented are maximum intensity projections of a z-stack (z = 0.3 μM). Nuclear size was analyzed with volocity software (version 6.3.1).