Identification of mRNAs that are spliced but not exported to the cytoplasm in the absence of THOC5 in mouse embryo fibroblasts

Depletion of the THOC5 gene in fibroblasts down-regulates cell growth. (A) MEF THOC5 flox/flox were infected with adenovirus carrying the GFP gene (Ad-GFP) or adenovirus carrying the GFP and Cre-recombinase genes (Ad-GFP-Cre) (Vector Biolab). Cell extracts were then examined by THOC5- and GAPDH-specific immunoblot. (B) RNAs were isolated from cells treated as above and were then analyzed by RT-PCR using actin- and THOC5-specific primers: forward primer: 5′-tctgccttttcacctggaag-3′; reverse primer: 5′-ctcggtacttttctgccagc-3′ (product size = 643 bp [wild type] or 429 bp [without exon IV/V]). (C,D) 10⁴ MEF THOC5 flox/flox or mouse NIH3T3 cells were infected with Ad-GFP or Ad-GFP-Cre virus, and cells were counted each day. Results represent the average values ±SD from three independent experiments.