Capture RIC-seq reveals positional rules of PTBP1-associated RNA loops in splicing regulation. Ye et al.
Original data of gels and blots for the manuscript "Capture RIC-seq reveals positional rules of PTBP1-associated RNA loops in splicing regulation"
Steps to reproduce
For RT-PCR, RNA was extracted with TRIzol and then treated with RNase-free DNase to remove genomic DNA contamination. Next, the resultant RNA was reverse transcribed with random hexamer primers before PCR. PCR products were run on 2% agarose gel and stained with Ethidium Bromide. For co-IP, cells were lysed with NP40 lysis buffer (10 mM Tris-Cl, pH 7.4, 100 mM NaCl, 2.5 mM MgCl2, 0.5% NP 40) containing proteinase inhibitors. Target proteins were pulled down with protein A/G beads coated with specific antibody and then eluted from the beads with LDS sample buffer and fractionated on SDS-polyacrylamide gels. To prepare samples for western blot, cells were lysed in 1× SDS sample buffer (50 mM Tris-HCl, pH 6.8, 2% SDS, 10% glycerol) and fractionated on SDS-polyacrylamide gels.