Figure 2.
Rpl5 intronic mutation identified in Skax23m1Jus mice leads to decreased mRNA, protein levels, and polysome defects. (A) Chromatogram of reverse-strand partial sequences of exon 1 and intron 1 of Rpl5 showing the heterozygous mutation (triangle) at the sixth nucleotide in intron 1. The exon-intron junction is indicated by the dotted line. (B) RT-qPCR showed a ∼50% reduction in Rpl5 mRNA in the Rpl5+/− mice when compared with WT ** P< .01. (C) Western blot analysis with vinculin and RPS19 used as loading controls showed reduced levels of RPL5 protein in Rpl5+/− mice when compared with WT. (D) Results from 3 separate experiments indicate Rpl5+/− mice have reduced 60S subunits, 80S ribosomes, and polysomes, consistent with Rpl5 haploinsufficiency.