z********i
发帖数: 610 | 1 Thanks very much for your reply.
Although more info regarding your experiments is needed, but basically,
1. it is possible that WT hes1 activated promoter but dnHes1 (dominant
negative?) did not; it only suggests that the endogenous Hes1 in the cell
line you tested was not invoved in maintaining the basal activity of that
promoter. Actually, it is not a bad observation; it suggests that hes1
activated this promoter via DNA binding, becaseu usually dnHes should have
no or little DNA binding activ... 阅读全帖 |
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r*******y
发帖数: 48 | 2 1. 我没有做的很细致，但是我看到了dnHes1能antagonize WT Hes1 induced promoter
activity.
That is good;
2. 保守的序列我已经做了突变，但是仍然能看到WT Hes1的激活作用，因为有好几个
potential Hes1 binding sites，所以我不能排除其它的位点不起作用。我现在在做
truncation mutation，想找到Hes1 responsive element。
how long of this promoter you tested? mutation of that conserved binding
site resulted in how much percentage of reduction in its activation compared
with that of the WT promoter by Hes1? in most cases,it w ill not be only
one site that contributes 100% to that activation... 阅读全帖 |
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r*******y
发帖数: 48 | 3 Although more info regarding your experiments is needed, but basically,
1. it is possible that WT hes1 activated promoter but dnHes1 (dominant
negative?) did not; it only suggests that the endogenous Hes1 in the cell
line you tested was not invoved in maintaining the basal activity of that
promoter. Actually, it is not a bad observation; it suggests that hes1
activated this promoter via DNA binding, becaseu usually dnHes should have
no or little DNA binding activity.
2. However, if you test both... 阅读全帖 |
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z********i
发帖数: 610 | 4 通过array找到了一个Notch的target gene，在细胞里面验证了，NICD下调，dnMAML上
调，我们猜想Hes1参与了调控，Hes1 cell line也验证了一下。然后克隆了target
gene promoter，预测发现了一个非常保守的Hes1 binding site，但是这个结果是WT
Hes1上调promoter，dnHes1不调节promoter activity，非常奇怪的结果。
想问一下Notch方面的专家，Hes1能直接上调gene 表达吗？
多谢了 |
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w*e
发帖数: 740 | 5 大家都用那家公司的HES1 AND HES5 做免疫荧光冰冻切片染色？
我买了几个都不WORK（虽然有的文章用过的），所以来咨询一下大家，谢谢 |
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g**********9
发帖数: 24 | 6 路过.
我刚开始做HES1 HEY1,希望有机会与你讨论问题! |
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z********i
发帖数: 610 | 7 Thanks so much for your suggestions.
刚才重新看了一下data，发现有一些变化，我以前忽视了。potential Hes1 binding
sites 突变之后，induction fold 重两倍变化了5倍变化为3.7倍。我在试试看是否能
的到稳定的结果。 |
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z***d
发帖数: 131 | 8 挺忙没时间审.有意要推荐的，站内留名,简要经历及e-mail address.
Keyword: Notch, Hes1, leukemia.
Journal IF: 1+. |
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z***d
发帖数: 131 | 9 【 以下文字转载自 Immigration 讨论区 】
发信人: zymed (abcg), 信区: Immigration
标 题: Leukemia审稿机会
发信站: BBS 未名空间站 (Wed Feb 25 19:21:34 2015, 美东)
挺忙没时间审.有意要推荐的，站内留名,简要经历及e-mail address.
Keyword: Notch, Hes1, leukemia.
Journal IF: 1+. |
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