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. 2013 Nov;19(11):1575–1582. doi: 10.1261/rna.041665.113

FIGURE 1.

FIGURE 1.

xCR1 mRNA translation can be repressed by Bicaudal-C. (A) Diagram of repression assay. Luciferase reporter mRNAs Luc/xCR1 (contains the 3′ UTR of the xCR1 mRNA) or Luc/Cyclin-BI (contains the 3′ UTR of the cyclinB1 mRNA) were injected into Xenopus embryos with or without the mRNA encoding candidate repressors (HA-Bic-C or MYC-Nanos1). Injected embryos were assayed for luciferase (Sheets et al. 1994; Fritz and Sheets 2001; Zhang et al. 2009), and the ratio of luciferase with and without a putative repressor was calculated as a measure of repression. (B) Bic-C specifically repressed the Luc/xCR1 reporter mRNA. (C,D) Regions of the xCR1 3′ UTR used in luciferase reporter mRNAs. The TCE (translational control element) was previously referred to as the Mut2 region (Zhang et al. 2009) and shown to be sufficient for repression in vegetal cells. Reporter mRNAs were analyzed for repression by Bic-C as described in A and B. (E) Repression by Bicaudal-C required a 5′ cap. The CSFV-Luc/xCR1-TCE reporter mRNAs contain the IRES from the CSFV 5′ of the luciferase coding region and the TCE of the xCR1 3′ UTR. The reporter mRNAs were analyzed for repression by Bic-C as described in A and B.