Figure 6.

Co-expression of catalytically active CLP1 with a tricRNA reporter in HEK cells reduces tricRNA formation. HEK 293 cells were transfected with a human tRNA-Tyr reporter containing Broccoli in the intron. Co-expression was conducted using the TSEN proteins and wildtype or kinase dead CLP1. Total RNA was isolated from HEK cells. (A) RNA was run on TBE-Urea gels and visualized for Broccoli containing RNAs using DFHBI-1T. Gels were washed and re-staned with SYBR Gold to visualize total RNA, which is shown as a loading control. (B) For the co-expression experiments involving wildtype and kinase dead CLP1 with the tricRNA reporter and the TSEN proteins, the respective tricBroc band was normalized to a band from the SYBR Gold (see ‘*’ in A) and quantified. The average and standard error for two biological replicates and a technical replicate are shown, a student's t-test was performed. (*P < 0.05).