GSM7900806: patient-derived fibroblasts, C9 NRE+ cytoplasmic ASO captured, FB506; Homo sapiens; RNA-Seq
Fibroblasts were first subcellular fractionated. Total RNAs were extracted from each fraction by using TRIzol LS reagent (ThermoFisher) according to the manufacturer's instructions. Residual DNA in both fractions was removed from RNA extracts by using TURBO DNA-free (ThermoFisher). NRE-RAP was performed similarly as the previously described RAP method (Engreitz, J. M. et al., 2014) with 5ʹ biotinylated (CCCCGG)3 ASO. The final RNA samples were cleaned up by using Monarch RNA Cleanup Kit (New England Biolabs). After ribosomal RNA depletion (Takara, 634846), RNA libraries were prepared using SMARTer Pico-Input stranded RNA-seq kit (Takara, 634836). For input RNA samples, RNA-seq libraries were constructed using a KAPA RNA HyperPrep Kit with RiboErase (Roche).