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Apoptotic cells were detected after the sections were incubated in the 3,3-diaminobenzidine (DAB) chromogen for approximately 6 min and counterstained with 4,6-diamidino-2-phenylindole (DAPI)

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All qRT-PCR samples were run in technical duplicates and the Ct method based on the reference value estimate from 3 housekeeping gene (GAPDH, GUSB and TBP) was used to calculate relative expression level of each gene and reaction