Fig. 5

Ropivacaine interacts with AKT1 directly to repress NF-κB signaling. A The 2D and 3D structure of ropivacaine hydrochloride and the predicted binding modes of ropivacaine hydrochloride with AKT1 kinase domain. B-C The stabilizing effect of ropivacaine on AKT1 protein was assessed by western blot in cellular thermal shift assay using the indicated concentrations of ropivacaine-treated MCF-7 and MDA-MB-231 cells. D The effect of ropivacaine on the activity of AKT1 kinase has been estimated through kinase activity assay. E The competitive binding relationship between ropivacaine and ATP was confirmed by using a pull-down assay. F Protein levels of p-AKT1, t-AKT1, p-iκBa, iκBa, p-NF-κB and NF-κB in the indicated concentrations of ropivacaine-treated MCF-7 and MDA-MB-231 cells were determined by western blot. Results are shown are shown as mean ± S.D from at least three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 (unpaired two-tailed Student’s t test)