Fig. 7: SLF2 and SMC5 patient cells exhibit mosaic variegated hyperploidy, mitotic abnormalities and sister chromatid cohesion defects.

a Quantification of the numbers of chromosomes per metaphase in peripheral blood lymphocytes from SLF2 or SMC5 patients, or an unrelated WT individual. 200 metaphases were counted in total from 2 independent blood samples. b Average number of mitotic cells with mis-segregated lagging chromosomes in SLF2 and SMC5 mutant fibroblast cell lines infected with lentiviruses encoding WT SLF2, WT SMC5, or an empty vector. n = 3 independent experiments for SLF2-P1, SMC5-P7 and SMC5-P8, and n = 4 independent experiments for SLF2-P2. A minimum of 250 mitotic cells were counted. c Representative images of mitotic cells from (b) with lagging chromosomes (scale bar: 10 µM). d Average number of mitotic cells with mis-segregated lagging chromosomes in U-2 OS SLF2 CRISPR HM cells infected with lentiviruses encoding WT SLF2 or an empty vector. n = 3 independent experiments. A minimum of 190 mitotic cells were counted. e Left: percentage of metaphases with rail-road chromosomes in peripheral blood lymphocytes from SLF2 or SMC5 patients, or an unrelated WT individual. A minimum of 380 metaphases were counted in total from 2 independent blood samples. Right: Representative images of metaphases (scale bar: 10 µM). f Percentage of metaphases with premature chromatid separation following 4 h treatment with 25 μM MG132 in SLF2 and SMC5 patient LCLs. n = 4 independent experiments. 200 total metaphases were counted. g Percentage of S/G2 cells (CENPF positive cells) with >2 centrosomes with or without 24 h exposure to 250 nM APH. n = 3 independent experiments. A minimum of 900 CENPF positive cells were counted. h Percentage of mitotic cells in SLF2 and SMC5 mutant LCLs with multi-polar spindles in untreated cells and cells exposed to 250 nM APH for 24 h. A minimum of 300 mitotic cells were counted over 3 independent experiments. i The percentage of G1-phase cells (CENPF negative cells) with >5 53BP1 bodies in SLF2 and SMC5 mutant fibroblast cell lines, with or without 24 h exposure to 500 nM APH. n = 4 independent experiments. A minimum of 390 G1-phase cells were counted. In all cases, a Student’s t test (two-sided, equal variance) statistical test was performed and error bars denote SEM.