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Paraffin blocks were sectioned onto SuperFrost Plus slides, dried overnight, and baked for 1 h at 60 C before immediately proceeding with the RNAScope Multiplex Fluorescent v2 assay according to the manufacturers instructions with the following modifications: Target retrieval was performed for 15 min in a boiling beaker, Protease III was used for 30 min pre-treatment at 40 C and hybridized slides were left overnight in 5 SSC before proceeding with the amplification and labeling steps

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2011 , (CRC Press, Taylor & Francis Group, 2011)