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qRT–PCR of BrU-labeled transcripts (Supplemental Fig. S2F)
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Cells were labeled with 2 mM bromouridine for 30 min, RNA was purified with Trizol, and labeled RNA was isolated by immunoprecipitation with B44 monoclonal anti-BrdU as described (Paulsen et al. 2014). Random primed cDNA was made with SuperScript III (Invitrogen), and qPCR was performed with the primers listed in Supplemental Table S2. Fold change was calculated using the Δ/ΔCt method.
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