Cells were pre-incubated with JTE-013 (10 µM) for 30 minutes and then treated with individual conjugated bile acids (TCA, TDCA, TUDCA, GCA, GDCA, 50 µM) or S1P (100 nM) for 30 minutes at 37°C. The protein levels of phosphorylated ERK1/2 (p-ERK1/2) and AKT (p-AKT) were determined by Western blot analysis and normalized with total ERK1/2 (T-ERK1/2) and total AKT (T-AKT) as described in Materials and Methods. The representative Western blot images are shown. The relative density was determined by Image J software.