Figure 10.

Number of dysmorphic nuclei with clustered NPCs increased with replicative senescence. (a) Representative images of cells stained with α-p16 and α-POM121 antibodies, counterstained with DAPI. White arrows indicate dysmorphic nuclei with clustered NPCs and elevated p16. (b) The numbers of dysmorphic nuclei with clustered NPCs, in cultures ≤5% senescence and ≥30% senescence of control and HGPS cells, were determined by counting dysmorphic nuclei positive for clustered NPCs and p16 positive signal, counterstained with DAPI. Differences between young and old cells in each cell line are significant. (c) The numbers of dysmorphic nuclei with clustered NPCs and progerin positive. On average, 87% of dysmorphic HGPS nuclei with clustered NPCs are positive for progerin. No significant difference could be determined depending on replicative senescence. (d) Representative images of HGADFN003 stained with α-NUP107 and α-progerin antibodies, counterstained with DAPI. White arrows indicate NUP107 clusters overlapping with strong progerin signal. At 5% senescence, HGPS cell passages were ≤P18, and control cells were ≤P21. At 30% senescence control passages were ≥P25, HGPS were ≥P21. Values are presented as mean ± SD (n ≥ 3), ns p > 0.05, *** p < 001, (b) two-way ANOVA with Tukey’s multiple comparison test, (c) one-way ANOVA with Sidak’s multiple comparison test.