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The cDNA was then fourfold diluted and used as a template in quantitative PCR with IQ SYBR Green Supermix (Bio-Rad) on a CFX96 instrument (Bio-Rad) using a 3-step protocol

After washing, cells then were mounted with Prolong Diamond Antifade Mountant containing 4,6-diamidino-2-phenylindole (DAPI) (Invitrogen) on glass slides