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All 10 carriers with a strong family history also had a morphology score of 5 or more, and these carriers comprised 48% of all probands with a strong family history and a morphology score of 5 or more.
The best culture (score = 7) in M2 also showed a medium TEER output (Fig. 2b); the only outlier in M3, which had a low morphology score of 5 (confluence was given, see S1), exhibited a low TEER reading (Fig. 2c).
These are the cancers, illustrated in Figure 1A, that have a morphology score of 5 or more but, despite extensive testing, are from women for whom we have not been able to identify germline BRCA1 mutations.
Overall, both charcoal-stripped media lead to poor morphological quality: Cells were dedifferentiated, flat or dwarf, lost cilia and secretory protrusions (Fig. 1a, b), with an average morphology score of 2.6 in M1 and 4.2 in M2 (Fig. 2a, b).
Figure 1A shows that mutation carriers tended to have higher morphology scores than non-carriers; of the carriers, 27 (93%) had a morphology score of 5 or more compared with 25% of all probands.
No diploid strains achieved a complex colony morphology score of 5, a complex mat score of 5, or a flocculation score of more than 0.3, where the highest haploid flocculation score was 0.8 (strain UWOPS83-787.3) and 16 strains achieved a score of 0.3 or higher.
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There is a striking relationship between the morphology score and differential expression of genes.
Expression patterns of genes that correlate with morphology are similar among normal to moderately deformed embryos (1 3 on morphology score scale), while severely deformed embryos (score of 4) show different patterns of gene expression.
Figure 1B shows that, when restricted to probands with a strong family history, the distribution of the morphology score was bimodal.
The expression pattern of genes for each morphology score group is presented in Figure 3b.
We also used residuals for a gene-by-gene analysis of morphology: rijk = μ + Ai + Dj + Mk + εijk where Mk is the kth morphology (morphology score 1 – 5 where 1 is normal and 5 is extremely deformed).
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