Decreased DNA density is a better indicator of a nuclear bleb than lamin B loss.

Abstract

Nuclear blebs are herniations of the nucleus that occur in diseased nuclei and cause nuclear rupture leading to cellular dysfunction. Chromatin and lamins are two of the major structural components of the nucleus that maintain its shape and function, but their relative roles in nuclear blebbing remain elusive. To determine the composition of nuclear blebs, we compared the immunofluorescence intensity of DNA and lamin B in the main nucleus body to that in the nuclear bleb across cell types and perturbations. DNA density in the nuclear bleb was consistently decreased to about half that of the nuclear body whereas lamin B levels in the nuclear bleb varied widely. Partial wave spectroscopic (PWS) microscopy recapitulated the significantly decreased likelihood of high-density domains in the nuclear bleb versus body, and that it was independent of lamin B level. Time-lapse imaging into immunofluorescence revealed that decreased DNA density marked all nuclear blebs whereas decreased lamin B1 levels only occurred in blebs that had recently ruptured. Thus, decreased DNA density is a better marker of a nuclear bleb than lamin B level.