私は大丈夫だわ。ああいう散漫な人たちとは違うから。はい。
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(1) Figure 1a and b in the Letter both show embryos generated from STAP cells, not a comparison of ES- and STAP-derived chimaeric embryos, as indicated in the legend.
小保方さんはaはESキメラだと思っていたんだけど、それだけではないわね。GFPはRosa26-GFPだわね。
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We became particularly interested in this question after a blastocyst injection assay revealed an unexpected finding. In general, progeny of injected ES cells are found in the embryonic portion of the chimaera, but rarely in the placental portion (Fig. 1a; shown with Rosa26-GFP). Surprisingly, injected STAP cells contributed not only to the embryo but also to the placenta and fetal membranes (Fig. 1b and Extended Data Fig. 1a–c) in 60% of the chimaeric embryos (Fig. 1c).
小保方さんはAC-129の実験がそれだと思っていて、かつ、そのESとそのキメラもあると思っていた可能性があるのよね。
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We tested the following three different genetic backgrounds of mice for STAP stem-cell establishment from STAP cell clusters, and observed reproducible data of establishment: C57BL/6 carrying Oct4-gfp (29 of 29), 129/Sv carrying Rosa26-gfp (2 of 2) and 129/Sv × C57BL/6 carrying cag-gfp (12 of 16). STAP stem cells with all these genetic backgrounds showed chimaera-forming activity.
FLSの樹立実験とAC-129の実験の前に、小保方さんにRosa26-gfpだと思いこませる
言い方で渡されたマウスがあるということだね。それが一応唯一胎盤にGFPの免染確認されている
Extended Data Figure1ということになるんだけど、その背景は
B6GFP × 129/Sv mice (B6GFP, C57BL/6 line with cag-gfp transgene)だね。
このマウスが理研に無いと言ったのは桂報告なんだけど、先にも見たように
丹羽さんは検証実験で使う予定で準備してるよね。
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R26R-H2B-EGFP transgenic mouse (Rosa-GFP) line was generated by Laboratory for Animal Resources and Genetic Engineering (LARGE), RIKEN CDB.
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We became particularly interested in this question after a blastocyst injection assay revealed an unexpected finding. In general, progeny of injected ES cells are found in the embryonic portion of the chimaera, but rarely in the placental portion (Fig. 1a; shown with Rosa26-GFP). Surprisingly, injected STAP cells contributed not only to the embryo but also to the placenta and fetal membranes (Fig. 1b and Extended Data Fig. 1a–c) in 60% of the chimaeric embryos (Fig. 1c).
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We tested the following three different genetic backgrounds of mice for STAP stem-cell establishment from STAP cell clusters, and observed reproducible data of establishment: C57BL/6 carrying Oct4-gfp (29 of 29), 129/Sv carrying Rosa26-gfp (2 of 2) and 129/Sv × C57BL/6 carrying cag-gfp (12 of 16). STAP stem cells with all these genetic backgrounds showed chimaera-forming activity.
Breaking News: Growing Concerns Over STAP Cell Sources
＞ However, I did not make chimera from those cell lines. Then I left RIken.
" In another email he clarified: "I left Riken one year ago. Then we could
not (establish) contact well between authors. Maybe Dr. Sasai, who wrote
the papers, thought that I did chimera experiment of 129.”
A troublesome part of the paper is the account of chimaera production. The details are very confusing and in part contradictory. Most glaringly we are told in the legend for Figure 5b that the Oct4-GFP transgene is used to show wide tissue contribution. I assume this is an error and a constitutive GFP reporter was used, but as pointed out above, at no point in the manuscript is there a description of how STAP cells are Isolated without use of the Oct4 reporter.
Natureの査読の
I assume this is an error and a constitutive GFP reporter was used
の部分から、CAG-GFPの記載はリバイス後らしい
