20211216文献精读{基因+通路}：功能回复实验

二维码

Silencing of ITGB6 suppressed CC progression via regulating JAK-STAT3 signaling.


<hr/> (A) Western blotting assay was applied to detect the JAK/STAT3 signaling pathway-related proteins (JAK1, p-JAK1, JAK2, p-JAK2, p-STAT3, and STAT3) in Hela and SiHa cells


<hr/>(B) The cell proliferation of Hela and SiHa cells was evaluated by colony formation (crystal violet stain, magnification: 40×)


<hr/>(C,D) Transwell assay was used to evaluate the migration and invasion of Hela and SiHa cells (crystal violet stain, magnification: 40×)


<hr/> (E) Western blotting assay was applied to detect the EMT-related proteins (Snail, vimentin, N-cadherin, and E-cadherin) in Hela and SiHa cells.


<hr/>结果描述


<hr/>这张figure是这篇文章的核心数据，实验都是常规实验，主要注意的是实验分组。这是典型的rescue实验分组。
rescue实验是机制研究的必备操作，如何设计，咱们根据一张模式图去理解。
酸性微环境诱导激活转录因子FOXX1高表达激活下游靶分子MAZ高表达，从而促进自噬阻碍了EMT过程。


<hr/>A单因素挽救

酸性微环境过表达FOXK1促进转移。在酸性微环境中使用RNAi抑制FOXK1观察转移能力是否下降。
B双因素挽救

FOXK1抑制自噬，自噬阻碍EMT。
低表达FOXK1同时加入自噬抑制剂观察EMT。
或者高表达FOXK1同时加入自噬诱导剂观察表型。


<hr/>值得注意的是，双因素的两个因素之间的调控关系可以是促进也可以是抑制。促进的比如说转录因子促进靶基因表达，抑制的比如说microRNA抑制mRNA。但是，调控关系一定是相反的操作才有意义。

原文地址：https://zhuanlan.zhihu.com/p/445915099