Figure 6 | Scientific Reports

Figure 6

From: Y-box protein-associated acidic protein (YBAP1/C1QBP) affects the localization and cytoplasmic functions of YB-1

Figure 6

YBAP1 abrogates cytoplasmic functions of YB-1. (a) An experimental scheme to examine mRNA stability using a d2EGFP reporter mRNA in HeLa Tet-Off cells. (b) Cells were transfected with pTRE-d2EGFP-AU, pEGFP2C1, pCMV-YB-1-3xFLAG and pCI-neo-YBAP1. Transcription from the Tet-regulated promoter was induced for 4 h, and shut off by the addition of Tet. The cells were harvested at the indicated time points. d2EGFP mRNA and 2xEGFP mRNA as an internal control were detected by Northern blotting. (c) d2EGFP mRNA remaining at various time points after the Tet addition was quantified. Relative half-life of d2EGFP mRNA is shown. *P < 0.05. (d) An experimental scheme to examine the effect of YB-1 and YBAP1 on the mRNA binding of eIF4E. Purple ovals represent PABP. Light blue, green, and orange circles represent other mRNP proteins. (e) Luc mRNA (lane 1) or Luc-MS2 mRNA (lanes 2–10) was incubated with nuclease-treated RRL, YB-1-His, YBAP1-His, and GST-MS2. The mixture was subjected to GST-pulldown assays as described in Methods. The eluted proteins were then analyzed by immunoblotting.

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