Extended Data Fig. 1: Progerin induces cellular senescence and mitosis defects.
a. Western blot analysis of CyclinA2, H3K9me3 and progerin in progerin-expressing IMR90 cells. n = 3 biologically independent replicates. b. β-Gal staining analysis and quantification of SA-β-gal positive cells (%) in progerin-expressing IMR90 cells. n = 3 biologically independent replicates. Scale bars: 100 μm. c,d. Immunofluorescence analysis of Ki67 in progerin-expressing IMR90 cells. Representative images (c) and quantification of the Ki67 positive cells (%) (d). n = 3 biologically independent replicates. Scale bars: 10 μm. e,f. Immunofluorescence analysis of binucleated cells in progerin-expressing IMR90 cells. Representative images (e) and quantification of binucleated cells (%) (f). n = 3 biologically independent replicates. Scale bars: 10 μm. g,h. Immunofluorescence analysis of binucleated cells in CRL-1474 and HGADFN003 cells. Representative images (g) and quantification of binucleated cells (%) (h). n = 3 biologically independent replicates. Scale bars: 30 μm. i,j. Metaphase chromosome spread assay in progerin-expressing IMR90 cells. Representative images (i) and quantification of anueuploid cells (%) (j). n = 3 biologically independent replicates. k,l. Metaphase chromosome spread assay in CRL-1474 and HGADFN003 cells. Representative images (k) and quantification of aneuploid cells (%) (l). n = 3 biologically independent replicates. m,n. Immunofluorescence analysis of chromosome laggings in progerin-expressing H1299 cells. Representative images (m) and quantification of chromosome laggings cells (%) (n). n = 3 biologically independent replicates. Scale bars: 10 μm. o,p. Immunofluorescence analysis of spindles in progerin-expressing H1299 cells. Representative images (o) and quantification of abnormal spindles cells (%) (p). n = 3 biologically independent replicates. Scale bars: 10 μm. For bar and line graphs, data are presented as mean ± SD. Statistical significance was determined in b, d, f, h, j, l, n and p using two-tailed unpaired Student’s t-test.
