Hes6 binds to Twist1 and maintains the stability of Twist1, promoting epithelial-mesenchymal transition (EMT) of GC cells (IMAGE)
Caption
(A) The network diagram of potential Hes6 binding proteins from the BioGRID database. (B) The Hes6-Twist1 protein interaction pattern constructed by computer simulation and molecular docking, and the prediction of potential sites on the interface between the two molecules. (C) The interaction between Hes6 and Twist1 in AGS and SGC 7901 cells was investigated using an exogenous immunoprecipitation assay (exogenous Hes6 vector carries a Flag tag). (D–G) The confocal images showing colocalization of Hes6 (green) and Twist1 (red) in AGS and SGC 7901. Nuclei were counterstained with DAPI (blue). The protein fluorescence intensity quantification curve is shown on the right. (H) Western blotting assay of Hes6-related Twist1 protein stability. The protein translation blocker cycloheximide (CHX) (10 μM) was used to treat vector control and Hes6 overexpression groups of AGS and SGC 7901 cells, and the rate of Twist1 protein degradation was detected by Western blotting. (I, J) Relative degradation curves of Twist1 protein in different treatment groups. (K) The expression levels of EMT-related marker proteins in AGS and SGC7901 cells stably transfected with vector and Hes6 were detected by Western blotting. (L, M) Relative quantitative statistics of EMT-related marker protein bands. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001.
Credit
Can Fang, Zhiwei Peng, Yaru Sang, Zihao Ren, Kang Wang, Nuo Xu, Ying Li, Tingting Guo, Yinan Zhu, Shangxue Yan, Kongwang Hu
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