1.45 na oil immersion objective

These were performed exactly as described previously (Selyunin and Mukhopadhyay, 2015 (link)). To summarize details of the microscope in this study, we used a sweptfield confocal microscope equipped with a four-line high-power laser launch and a 100× 1.45 NA oil immersion objective (Nikon). An iXon3 X3 DU897 electron-multiplying charge-coupled device camera (Andor Technology) was used for image capture. All images were captured as z stacks with 0.2-µm spacing between individual frames. Images depicted in the figures are maximum-intensity projections of the stacks.
All analyses were performed using ImageJ (National Institutes of Health) as described previously (Selyunin and Mukhopadhyay, 2015 (link)).

For immunofluorescence, images were collected on a Nikon TiE microscope equipped with a Yokogawa CSU-W1 spinning disk confocal unit (50-μm pinhole size), an Andor Borealis illumination unit, Andor ALC600 laser beam combiner (405 nm/488 nm), Andor IXON 888 Ultra EMCCD camera, and a Nikon 100×/1.45 NA oil immersion objective. The microscope was controlled by software from Nikon (NIS Elements, ver. 5.02.00). DAPI or Alexa 488 were excited with 405-nm and 488-nm lasers. Within each experiment, cells were imaged using the same settings on the microscope to compare signal intensities between cell lines. Fiji software was used to analyze images. The region of interest of the entire nucleus was manually selected or thresholded using the DAPI signal. Fluorescence intensities of these regions of interest were measured in the 488-nm channel. The mean fluorescence intensities were extracted for each cell line.