Extended Data Fig. 7: Nuclear pore defects in Grn^−/−neurons treated with Grn^−/− MCM.

a, 3D structured illumination microscopy (SIM) images of Nup98 and LMN A/B in Grn^+/+ and Grn^−/− cortical neurons treated with control media, Grn^+/+ MCM and Grn^−/− MCM (250 μg/ml). Nup98 is shown in red, Lamin A/B in green and MAP2 in blue. b, Nup98 intensity distribution per intranuclear grid (0.44 × 0.44 μm²) in Grn^+/+ and Grn^−/− cortical neurons treated with control media and Grn^−/− MCM (250 μg/ml) (see Methods for specific algorithms). Nup98 is less evenly distributed in Grn^−/− cortical neurons in control media. When Grn^+/+ neurons are treated with Grn^−/− MCM, they show significant uneven distribution of Nup98 compared to Grn^+/+ neurons treated with control media. Interestingly, Grn^−/− neurons treated with Grn^−/− MCM do not show further defects in Nup98 distribution compared to Grn^−/− in control media. Data represent mean ± s.e.m. from 3 independent cultures. Statistics use two-way ANOVA. c, Average of Nup98 intensity in the small grids in Grn^+/+ and Grn^−/− cortical neurons treated with control media, Grn^+/+ MCM and Grn^−/− MCM (250 μg/ml). Data represent mean ± s.e.m. The numbers listed below each data set represent the number of neurons analysed from 3 independent cultures. Statistics uses two-tailed, unpaired Student’s t-test.