Figure 1. Nuclear envelope rupture during interphase. (A) U2OS cells transiently transfected with GFP₃-NLS and imaged every 3 min for 36 h show transient interphase rupturing of the NE followed by recovery of GFP₃-NLS into the nucleus. (B) Dynamics of a rupture event. U2OS cells expressing GFP-NLS were imaged every 30s to capture NERDI in high temporal resolution. GFP intensity was normalized by setting the maximum and minimum intensity for each cell to 1 and 0, respectively. Curve fittings of individual interphase NE ruptures were plotted (lines) along with raw data (points). Data was fit using the equation: Y = IF(X < X0,Y0,IF(X < X1,Y0-S*X,Bottom+(Top-Bottom)/(1+10^((Log50-X)*HillSlope)))) where: X0 is the point of inflection between the plateau and the spilling event, Y0 is the plateau value, X1 is the initial point of recovery, S is the slope of spilling, Bottom is the lower plateau for recovery, Top is the upper plateau of recovery, Log50 is the point of 50% recovery, and HillSlope is the linear rate of recovery. (C) Representative images of HeLa cervical and SJSA osteosarcoma cancer cell lines demonstrating spilling in diverse cancer cell types. (D) U2OS cells transiently transfected with GFP₃-NLS and imaged every 3 min show localized nuclear deformation and cytoplasmic GFP signal originating from the site of deformation.