Landscape of Star-PAP mRNA association along with its co-regulator RBM10 using HITS-CLIP (high-throughput sequencing coupled with cross-linking immunoprecipitation).

We employed HITS-CLIP to map genome wide Star-PAP mRNA binding and define the role of RBM10 on global Star-PAP mRNA association. We show a transcriptome-wide association of Star-PAP which is diminished on cellular Star-PAP depletion. HITs-CLIP data analysis of RBM10 knockdown and pulldown with anti Star-PAP antibody on HEK293 cells indicates significance of RBM10 in Star-PAP and mRNA association. Star-PAP HITS-CLIP was performed in HEK- 293 cells after transient knockdown of either RBM10 or Star- PAP. RNA was pulled down using anti-StarPAP antibody or anti-IgG as mentioned in the protocol.