Lamins are removed during nuclear shrinking in a PKC-dependent manner. The nuclear shrinking assay was performed as in Fig. 1. Z-stack images were acquired by confocal microscopy. For intensity measurements, all images were acquired with the same exposure time. NPC and LB3 density at the NE were determined by quantifying staining intensity within a fixed defined area (20 µm2) for an individual z slice at the periphery of the nucleus. Total NE staining intensity per nucleus was estimated from the density measurement and total NE surface area. Shown in A and C are representative maximum-intensity projections and z slices at the periphery of nuclei used for intensity measurements. Bars, 5 µm. (A and B) NPCs were visualized with mAb414 (red). n > 10 different nuclei in two different extracts. (C and D) LB3 was visualized with an α-LB3 antibody (green). n > 10 different nuclei in two different extracts. ***, P < 0.005. Error bars represent SD.
