丹羽論文のここにRFPという言葉が出で来るんだけど、李がどういうつもりの嘘をついているのかというあたりだな。
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Figure 5: アnalyses of fluorescent signals from GOF transgene.
Figure 5
(a) Fluorescent microscopic analysis of cell aggregates derived from GOF Tg mice. The cell aggregates (top) were derived from liver cells of 6-days old of GOF Tg mice. Fluorescent images with the filter sets for detection of GFP and RFP signals are shown. Images of ES cells carrying CAG-GFP (middle) captured with the same conditions are shown as a control to confirm no leaky signal of GFP in RFP channel. Images of wild-type ES cells (bottom) are shown as a control to confirm spesific signal of GFP in GFP channel. (b) FACS analysis of the low-pH treated spleen cells derived from GOF Tg mice. The spleen cells were isolated from 7-day-old GOF Tg mice and prepared with Lympholyte followed by treatment with the indicated stressors. After the culture for seven days, the cells were dissociated, stained with anti-E-cadherin with PE and anti-CD45 with APC, and analyzed by FACS. Wild-type ES cells were used as a positive control for E-cadherin staining and a negative control for CD45-staining as well as GFP fluorescence.
確かにキメラの子供のジャームライントランスミッションは兄妹交配でできたらある。
しかし、Nature誌の査読Referee #2は
”Table 2 reveals that chimaeras were inter-crossed. Why? This is very unusual. The data in the table cannot be fully interpreted without information on the strain, genotype and gender of the STAP cells. ”
と質問しています。
このTable 2はExtended Data Fig. 7c と思われますが、兄弟交配はvery unusualなんですね。
そこで、methodsに
”To make germline transmission more efficient, we intercrossed chimaeras in some experiments.”
と説明を入れたのでしょう。
といったところですか。
それで、先に引用したのところの前に
”Because the number of CD45+ cells froma neonatal spleen was small, we mixed spleen cells from male and female mice for STAP cell conversion.”
の説明も加えられたのでしょう。
丹羽Protocolに出てくる
(iv) Cells from mice older than one week showed very poor reprogramming efficiency under the current protocol. Cells from male animals showed higher efficiency than those from female.
当時の若山さんがどの程度調べていたかは別として、取り合えず丹羽さんの計算を
挙げましょうかね。
>>However, when we used ATP instead of HCl, also based on a suggestion by the authors, a few cells in a subset of cell aggregates expressed the pluripotency marker Oct3/4 at levels comparable to those in ES cells that were reproducibly detected by QPCR (Fig. 3c) and immunostaining (Fig. 4b). However, the frequency was very low; 5 × 105 liver cells yielded only ~30 cell aggregates, in which about 20% of the cell aggregates contained 1–2 Oct3/4 positive cells, indicating a frequency per seeded liver cell of 0.0012–0.0024%.
丹羽さんはいわゆる政治的理由でそれを書いてないんだね。でも同じ条件ではないが
小保方さんがティシュー論文でそれをあきらかにしていたよね。
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Under a microscope, 2000 spheres, each containing *1000 cells, were collected using a glass pipette and placed in a 50mL tube.
(顕微鏡下で、それぞれ1000個の細胞を含んだ2000個の球状体をガラスピペットを用いて収集し、50mLチューブに入れた。)
