Reh cell line, input, replicate 1
RIP was performed according the protocol standardized previously in our lab. Briefly, Reh cells were lysed in a cocktail of Non-Denaturing Lysis Buffer and Protease plus RNAse Inhibitors. 10% of the precleared lysate was stored as Input. The precleared lysates were incubated overnight with Protein G Agarose beads along with antibody against human IGF2BP1 (MBL RN007P) or normal Rabbit IgG (CST 2729). Pelleted beads were washed thrice with PBS and resuspended in Non-Denaturing Buffer the next day. RNA was extracted using QIAGEN miRNeasy RNA isolation kit as per the manufacturers protocol. A part of the RNA samples was subjected to reverse transcription and qPCR and the rest was sent for RNA sequencing (Input RNA and IGF2BP1 immunoprecipitated RNA).