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A lower DNA methyltransferase activity increases expression of silenced genes such as TSGs reactivating expression of key genes.
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However, BS modified DNA sequencing showed that promoter methylation could not fully explain Clusterin basal and reactivated expression.
Importantly, however, after only 5 days EWS-FLI1 reactivated expression of 81 hNCSC-associated genes and inhibited expression of 126 hNC-MSC-associated genes (Table S3A).
To distinguish these possibilities, we isolated subclones from cells with silenced PTRE-HPRT that spontaneously reactivated expression and used selection for HPRT to maintain the reactivated state for at least one month (∼50 cell divisions).
The best evidence was the ability of TG-resistant cells to reactivate expression and restore functional HPRT activity, which was evident by growth of the cells in AzHx media.
Although inhibiting DNA methylation and histone deacetylation usually reactivates expression of silenced alleles [38], [39], such renewed expression is often unstable and quickly re-silences at a high frequency, possibly as a consequence of retention of some repressive histone modifications [29], [30].
Exposure to AZA reactivated expression in MAK-I and AK31, consistent with methylation-dependent silencing.
These demethylating agents have been shown to reactivate expression of numerous methylation-silenced genes.
Common to all Burkitt lymphomas, endemic or sporadic, are distinctive chromosomal translocations that reactivate expression of the c-myc protooncogene and comprise the primary oncogenic mechanism.
Mechanical loading clearly reactivated expression of the aggrecan and type II collagen genes at loading of 60 min/day compared to the other durations of loading.
The reactivated expression was subsequently ascribed to the E82 locus, rather than C73, by testing F2 segregants lacking C73 (see Figure 5 below).
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