n******w 发帖数: 70 | 1 没看后面的讨论,来尝试回答下问题
HYPOXIA和MET是有联系的,HYPOXIA在先,MET在后。可能机理如下
1,Hypoxia-HIF1a-VEGF/VPF-vascular permeability-tumor dissemination;
2,Hypoxia-HIF1a-VEGF-C-peri/intra-tumor infiltration of lymphatic vessel-
tumor dissemination.
3, Hypoxia-met niche-invasive front of tumor-tumor dissemination.
引用就不列了,PUBMED一下就出来。
Dissemination是时时刻刻都存在的,真正能否形成MET,就是一个几率问题。换言之,
DIS越多,MET约有可能形成。 |
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s******r 发帖数: 2876 | 4 Hif1a的promoter是不是厌氧的?
如题请推荐,另,有没有啥厌氧promoter?
3q |
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a*****u 发帖数: 802 | 5 proteasome substrates太多了,经典的有p53, cyclinB, HIF1a, HMG CoA reductase2
, Insig1 (忘了是1还是2)。
proteasome有3个active centers建议最直接的方法就是纯化proteasome,用产荧光的
substrates检测其活性。或者简单的用model substrates,其中一个是ub-GFP |
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r********s 发帖数: 149 | 6 I think everyone knows what hif1a means in the field. |
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A******y 发帖数: 2041 | 7 HIF1a is constantly degraded in normal cells. You won't even detect that
much nevertheless to see it decreases. Nrf2, however, is a completely
different story. |
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a*****r 发帖数: 54 | 8 胖老师,HIF field用的最多的HIF1A的抗体是BD的#610958(鼠单抗),WB的band在~
120kD
在normoxia培养条件的cell lysate信号几乎没有或者很低
需要进行一定的处理才能看见robust的信号,例如
(1)MG132 ~2hr
(2)CoCl2/iron chelator(比如DFO)
(3)PHD inhibitor(比如DMOG)
(4)如果你有Hypoxic incubator,用低于2% O2
(5)或是直接使用某些Mutant细胞,例如RCC4 (VHL inactivated) |
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