FIG. 7.

EIF4A3 and PRPF19 alter splicing of the Fshb and/or Lhb transcripts. A) SiRNA constructs targeting Eif4a3 or Prpf19 transcripts were transfected into LβT2 cells, and the respective protein levels assessed by Western analysis and quantified as in Figure 6C. B) The effect of these siRNA constructs on the Fshb and Lhb transcripts, as well as those of the targets, were measured by RT-PCR. C) The effect of overexpression of EIF4A3 on the Fshb transcript (representing spliced and unspliced forms, the primers amplify only exon 3, and do not span an intron) was measured similarly by RT-PCR in αT3–1 cells; the degree of overexpression was assessed by Western analysis (lower panel). D) The effect of activin (100 ng/ml, 24 h) on Eif4a3 mRNA levels and on the splicing of the Fshb primary transcript in LβT2 cells was assessed by RT-PCR using primers spanning the first intron, and relative levels of unspliced:spliced transcripts measured as in Figure 6A.