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The next day, sections were rinsed, incubated in secondary antibody in blocking solution for 2 h at room temperature on rotation and then rinsed and counterstained with Hoechst 33342 (1:2000 in PBS, Fisher #H3570) before being mounted on SuperFrost Plus slides (Fisher #12-550-15), dried and coverslipped with Prolong Gold Antifade mounting medium (Fisher #P36934)

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