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Slides were incubated in AlexaFluor 488-conjugated anti-rabbit secondary antibody (Thermo Fisher Scientific, Cat #A-11008) in 0.1 M PB containing 0.3% Triton X-100 and 1% donkey serum for 2 h at room temperature and immediately covered using Vectashield mounting media
The relative expression of target genes was calculated and normalized to that of hypoxanthine phosphoribosyltransferase 1 (HPRT) using the Delta C(T) method
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