三誌論文には幹細胞化の実験のことは書かれていないからね。まさに8月に提出する以前の
作業に関して語っているんだね。ここはどうなの?
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Letter Fig.2iは2012年にTruseqによるRNA-seqを行い、そのデータに基づいて作成し た系統樹と考えられるが、
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Letter Extended Data Fig.6dは、2012年にSMARTERキットによるRNA-seqを行い、その データをもとに作成した系統樹と考えられるが、
リジェンドは以下だね。
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3. Extended Data Figure 3: Transcriptome analyses of STAP cells shown by heat maps. (494 KB)
a, Heat maps of expression profiles of top-ranked up- and downregulated genes in STAP cells (Oct4-GFP+ clusters converted from CD45+ cells) compared to ES cells. Their respective expression levels in STAP stem cells, trophoblast stem cells and Fgf4-induced stem cells are shown. Absolute expression values are scaled by log2. The genes expressed differentially between ES cells and STAP cells tended to show more similar expression profiles to ES cells in STAP stem cells and Fgf4-induced stem cells than in trophoblast stem cells. Expression of some early endodermal lineage genes such as Gata4 and Sox17 was moderately elevated in STAP cells as compared to ES cells, whereas its biological significance remains elusive (these genes are shown to be strongly expressed in Oct4-GFP-dim cells1). b, Heat maps of expression profiles of top-ranked up- and downregulated genes in ES cells compared to CD45+ cells and their respective expression levels in STAP cells.
The genes expressed differentially between CD45+ and ES cells tended to show similar expression profiles in ES cells and STAP cells. c, Heat maps of expression profiles of representative genes implicated in haematopoietic lineage development in CD45+, ES and STAP cells. No strong correlation was seen between CD45+ cells and STAP cells in their expression profiles (a similar tendency of no correlation was seen for the data in b).
分かりにくいのはaとbとで遺伝子の並びが違うことだね。もう一度本文を見てみよう。
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To investigate the relationship among STAP cells, STAP stem cells, Fgf4-induced stem cells, ES cells and trophoblast stem cells, we performed genome-wide RNA-sequencing analysis (Fig. 2i for dendrogram; Extended Data Figs 3 and 4 for expression analyses of representative genes ; Supplementary Tables 2 and 3 for analysis conditions). Whereas STAP cells formed a cluster with STAP stem cells, Fgf4-induced stem cells, ES cells and trophoblast stem cells and not with the parental CD45+ cells, STAP cells were an outlier to the rest of the cell types in the cluster. In contrast, STAP stem cells were closely clustered with ES cells. Fgf4-induced stem cells formed a cluster with a sub-cluster of ES cells and STAP stem cells, whereas trophoblast stem cells comprised an outlier to this cluster, indicating a close relationship of Fgf4-induced stem cells with these pluripotent cells.
学さんの説明は端折りすぎだね。リジェンドは以下だ。
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j, qPCR analysis of Fgf4-induced cells (cultured under feeder-free conditions) with or without JAK inhibitor (JAKi) treatment for pluripotent marker genes.
k, qPCR analysis of FI-SCs with or without JAK inhibitor (JAKi) treatment for trophoblast marker genes. Values are shown as ratio to the expression level in ES cells (j) or trophoblast stem cells (k). ***P < 0.001; NS, not significant; t-test for each gene between groups with and without JAK inhibitor treatment. n = 3. Statistical significance was all the same with three pluripotency markers. None of the trophoblast marker genes showed statistical significance. Error bars represent s.d.
j,多能性マーカー遺伝子のためのJAK阻害剤(JAKi)処置の有無で分けられた(フィーダー無し条件下で培養された)Fgf4誘導細胞のqPCR分析。
k,栄養膜幹細胞マーカー遺伝子へのJAK阻害剤(JAKi)処置の有無で分けられたFI-SCsのqPCR分析。値はES細胞(j)または栄養膜幹細胞(k)の中の発現レベルに対する比として示されている。 *** P <0.001; NS、重要でない;JAK阻害剤処理の有無によるグループ間の各遺伝子についてのt検定。 N = 3。三つの多能性マーカーの統計的有意性はまったく同じであった。栄養膜幹細胞のマーカー遺伝子のいずれも統計的有意性を示さなかった。エラーバーは±標準偏差を表す。
親分、ここでしょ。
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Thus, our findings indicate that epigenetic fate determination of mammalian cells can be markedly converted in a context-dependent manner by strong environmental cues.