Figure 5.

USP39 is involved in Rictor pre-mRNA splicing and maturation. (A) Schematic illustrating the qPCR primer pairs used to measure pre-mRNA and mature mRNA species based on the Rictor gene sequence in the Ensembl database. (B, C) qRT-PCR analyses showing relative mRNA levels of spliced and unspliced Rictor RNA transcripts in ECA109 and KYSE30 cells with or without USP39 shRNA knockdown (B). Splicing efficiency is expressed as the ratio between mature/pre-mRNA levels (C). GAPDH was used for normalization. (D) RNA immunoprecipitations (RIP) performed using antibodies against USP39 or an IgG control. qRT-PCR were used to measure relative Rictor mRNA levels recovered in the immunoprecipitated complex. The results are shown as mean ± S.D. of three independent experiments. Student’s t-test: ^* p < 0.05, ^** p < 0.01, ^*** p < 0.001.