Figure 4.

Stable EJC Deposition Is Required to Block Recursive Splicing

(A) KD of EJC components was rescued with siRNA-resistant FLAG-tagged counterparts. The splicing pattern of endogenous KPNA1 or stably integrated aCADM2 RS-exons was monitored by RT-PCR.

(B) Wild-type or mutant SL2 sequence was inserted into KPNA1 and mCADM2 splicing reporters at the expected EJC deposition site, and the splicing pattern was monitored by RT-PCR after co-transfection of SL2 reporters and plasmids expressing the indicated MS2-tagged proteins.

A minimum of 3 independent replicates was performed in HeLa (A) or 293 (B) cells.