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Fig. 5 | BMC Biology

Fig. 5

From: Pathologic tau conformer ensembles induce dynamic, liquid-liquid phase separation events at the nuclear envelope

Fig. 5

Mislocalization of nucleoporins and nuclear membrane rupture in ES1 cells. a–d Nuclear deformation and mislocalization of nucleoporins. 4RD-YFP tau reporter (Ctrl) and ES1 cells were fixed. Lamin B1 (a) and nucleoporins (c) were probed and visualized with fluorescent conjugated secondary antibodies. Lamin B1 in magenta; nucleoporins in red; tau in green; nuclei were counterstained with hoechst 33342 (blue). Scale bar, 10 μm. b Morphometric shape descriptors, solidity and circularity, were measured along the nuclear margins (lamin B1 staining) to appraise nuclear deformations in ES1 line (n = 325) compared to controls (n = 245). Nuclei from 8 different areas (160 μm2 for each, see Supplementary Figure 5). d Colocalization of nucleoporins (using anti-nucleoporins antibody, NPC414) with tau inclusions were determined by measuring intensity of NPC414 signals within tau inclusions over 1 μm2 in size. Total 258 inclusions from 6 different areas (160 μm2 for each) were analyzed. The level of colocalization was presented as the frequency of tau inclusion per unit intensity of NPC414. a.u., arbitrary unit. e TEM analysis of control (left) (n = 2) and ES1 cells (right) (n = 6) showed a disruption of the double membrane architecture of the nuclear envelope (also see Supplementary Figure 6c). Arrowheads (blue, normal; red, ruptured) indicate nuclear envelopes. C, cytoplasm; N, nucleoplasm. Scale bar, 500 nm and 250 nm in the boxed images

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