Fig. 6: ERK2 directly binds and phosphorylates eIF4B Ser93 by which promotes mesenchymal markers translation. | Cell Death & Disease

Fig. 6: ERK2 directly binds and phosphorylates eIF4B Ser93 by which promotes mesenchymal markers translation.

From: EIF4B Ser93 phosphorylation by ERK2 promotes epithelial-mesenchymal transition to drive colorectal cancer metastasis

Fig. 6: ERK2 directly binds and phosphorylates eIF4B Ser93 by which promotes mesenchymal markers translation.

A The top six kinase which might regulate eIF4B Ser93 phosphorylation predicted by the “Kinase Prediction” website (https://www.phosphosite.org/kinaseLibraryAction) were shown. B The conserved amino acid sequence surrounding eIF4B S93 matches the consensus MAPK phosphorylation motif. C ERK2 was predicted as the kinase that regulated eIF4B Ser93 phosphorylation. D PPI network diagram of ERK2 (MAPK1) and eIF4B. E HT29 cell lysates were prepared by RIPA lysis and co-immunoprecipitated by ERK2 or eIF4B antibody with IgG as negative control. The immunoprecipitated protein was detected by western blotting. F GST-pull down assay showed the interaction between His-ERK2 and GST-eIF4B (T53-E145) protein in vitro. In vitro kinase assay and western blotting analysis of p-eIF4B (T53-E145)Ser93. The following assays were conducted in eIF4B WT and eIF4B S93D CRC cells following Vx-11e treatment (1 μM) or the control (DMSO) for 24 h. G The phosphorylation of eIF4B Ser93, eIF4B and EMT markers (Snai1, E-cad) protein were detected by western blotting. H The levels of mesenchymal makers mRNA were measured by RT‒qPCR. I RT-qPCR analysis of the mRNA abundance of mesenchymal markers in polyribosome fractions. All the data are presented as mean ± SEM from at least three independent experiments. P values were determined by two-sided Student t test. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

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