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RT-PCR using primers targeting exons 1 and 5 suggested that the top and bottom bands in the mutant cells represented the predicted transcript (with exon 1 joined to exon 4) and an aberrant transcript with exon 1 joined to exon 5, respectively, which was confirmed by restriction enzyme digestion of the amplicons (lanes 5 to 10).
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In previous experiments using pIREShyg2-EGFP- CA 13, the EGFP-positive population consisted of two distinct fractions, M1 and M2, which represent intermediate (correlating with replication errors) and permanent mutant cells (representing the combined effect of replication errors and MMR proficiency) (21).
It is worth noting that, as control and mutant cells represent only a proportion of the total cell number in the cocultures, this number is lower than the totals for each genotype when grown separately.
WT and mutant cells are represented as black and red histograms, respectively.
Thus, XP-mutant neural cells represent valuable tools to clarify the molecular mechanisms of neurological abnormalities in the XP patients.
Thus, chino-mutant T cells represent a mild deficiency in mTOR signaling reminiscent of a partial loss-of-function in mTOR (Zhang et al., 2011) that exclusively affects extrathymic CD4+ T cell differentiation as seen in conditional T-cell-deleted Frap1 knockout mice (Delgoffe et al., 2009).
B0 cells represent untreated cells.
These cells represent PBMCs.
Shaded cells represent significant p-values.
The presence of proper ploidy subsequently led us to hypothesize that the 'without errors' divisions that are present in ∼20% of esco2 mutant cells (Fig. 5B) represents those cells that displayed the PBS phenotype in chromosome spreads.
Tumors are considered as organ-like structures than just a clonal expansion of mutant cells, and their microenvironment represents an important issue for the development of new therapeutic strategies [ 24, 25].
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