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One microgram of total RNA was reverse-transcribed with a ReverTra Ace qPCR RT Kit (Toyobo, Japan) following the suppliers recommendations

Briefly, cells were lysed at 4 C in a homogenization buffer (210 mM mannitol, 70 mM sucrose, 5 mM HEPES, pH 7.4) supplemented with EGTA (Sigma-Aldrich), albumin 0.5% and protease and phosphatase inhibitors (Sigma-Aldrich) and centrifuged at 600 g for 3 min, 4 C to pellet nuclei and heavy particles

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Should supplemental antioxidant administration be avoided during chemotherapy and radiation therapy