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Thereafter, the TMB substrate was added, and the color was developed in the dark at 37 C for 15 min

High-LET radiation elicits qualitatively distinct biological responses at low versus high doses ( From DNA Damage to Stable Phenotype: The Role of the DDRcGASSTING Axis A critical question is how a transient physical insult (irradiation) induces stable, heritable changes in the cellular operating system. We propose that complex, clustered DNA double-strand breaks (DSBs) generated by high-LET heavy ions are the initiating trigger ( Beyond Epigenetic and Transcriptional Reprogramming: Genomic Scars as a Foundation for Phenotypic Stability Beyond epigenetic and transcriptional reprogramming, a fundamental mechanism driving the observed stable phenotypes may reside in the permanent genomic alterations elicited by high-LET radiation

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Briefly, hippocampi of newborn Wistar rat pups were dissected in HBSS (140 mM NaCl, 5 mM KCl, 6 mM glucose, 4 mM NaHCO 3 , 0.3 mM Na 2 HPO 4 and 0.4 mM KH 2 PO 4 ) and incubated for one hour in enzyme solution (DMEM containing 0.5 mg/mL cysteine, 100 mM CaCl 2 , 50 mM EDTA and 2.5 U/mL papain, bubbled with carbogen for 10 min)