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It is noteworthy that the PAR signal intensity did not increase further at 24 h.
In addition, different receptor crosstalk mechanisms critically contribute to a high diversity of PAR signal transduction and receptor-trafficking processes that result in multiple physiological effects.
In support of this idea, in our immunoprecipitation experiments we did not detect a PAR signal at the molecular weight of GFP-NEIL1, suggesting that NEIL1 is not significantly poly(ADP-ribosyl)ated in vivo (data not shown).
Thus, our interpretation of the effect of ATM inhibitor is that it promotes CHD4 accumulation indirectly by enhancing the PAR signal at DNA-damage sites (shown in Supplementary Figure S1F).
Typical for this posttranslational modification, the major PAR signal was detected as a high molecular weight smear, and most likely corresponds to auto(ADP-ribosyl)ated PARP (see Figure 4B).
In these reactions, we did not observe a shift in the mobility of GST-NEIL1, an indication of poly(ADP-ribosyl)ation, nor was there any detectable PAR signal around the molecular weight of GST-NEIL1, indicating that NEIL1 is not substantially poly(ADP-ribosyl)ated by PARP-1 (data not shown).
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This fits with the fact that PAR signals at DSBs are transient and disappear in 10 min.
At present, PAR signalling is known to activate several major signal pathways.
The function of Chd1l could be in PAR signaling via the PAR-binding macro domain and in downstream DNA repair.
The mechanisms whereby the NLRs can synergize with PAR signalling, as observed for oral pathogens [ 195], remain to be determined.
In addition, we discuss the suitability of these receptor interaction mechanisms as targets for modulating PAR signalling in disease.
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