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Cells were then treated with 100 mM reducing agent sodium 2-mercaptoethanesulfonate (Mesna) in TNE buffer (20 mM Tris, 150 mM NaCl, 1 mM EDTA, pH 8.6) or with TNE buffer alone for 5 minutes before being washed 1x in PBS and imaged in Hanks Balanced Salt Solution (HBSS) buffer on a Nikon Ti2E wide-field microscope with a light-emitting diode (LED) light source and a 20x/0.75 NA objective, with several phase contrast and epifluorescence images acquired per well using corresponding filters

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