Xenbase Image ID: 137433

Figure 3. An Exchange from Somatic to Oocyte Transcriptional Machinery Takes Place during Reprogramming by the Xenopus Oocyte(A) Confocal imaging of YFP-RPB1 expressing nuclei (yellow) transplanted into oocytes expressing cherry-H2B (magenta). Images were recorded 15 min and 15 hr after NT. Scale bar, 20 μm. Graph at right: average mean pixel intensity per nucleus normalized to the highest value.(B) GFP-TBP nuclei (green) transplanted into oocytes expressing TBP2-cherry (red). Confocal images were recorded soon after (15 min) and 24 hr after NT. Incorporation of TBP2-cherry and disappearance of GFP-TBP are observed.(C) Experimental set-up for western blot analysis and transcriptional inhibition with aAm or Flav.(D) Transcriptional inhibition by aAm and Flav inhibits activation of Oct4 and Sox2 in NT oocytes, as examined by RT-qPCR analyses. n = 3. Data are represented as mean ± SEM.(E) Western blot analysis of total Pol II, which recognizes both Pol IIA (hypophosphorylated) and Pol IIo (hyperphosphorylated), Ser5P Pol II, Ser2P Pol II, and histone H3 in MEF nuclei 0, 6, 24, and 48 hr after NT. Arrowheads indicate Pol IIA and Pol IIo. aAm and Flav treatments of transplanted nuclei are shown.(F) Comparison of the Ser2P Pol II band intensity (red) before and after NT. The band intensity, detected by western blot, was normalized to the number of nuclei. Fold enrichment of Ser2P Pol II in NT samples over donor cells is shown in the graph. Data are represented as mean ± SEM; n = 3.See also Figure S3 and Movies S1 and S2. Image published in: Jullien J et al. (2014) Image downloaded from an Open Access article in PubMed Central. © 2014 The Authors. The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society Larger Image Printer Friendly View

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