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. 2019 Nov 20;9:17152. doi: 10.1038/s41598-019-53676-3

Figure 4.

Figure 4

VIP induces glucose uptake through PKA, MAPK, PI3K and mTOR pathways in human trophoblast cells. BeWo cells were cultured as in Fig. 2. Cells were pre-incubated with the corresponding specific kinases inhibitors for 20 min before the addition of 50 nM VIP for 10 min and 2-NBDG for another 10 min. Cells were washed with cold PBS and flow cytometry was performed. (a) 2-NBDG uptake in absence/presence of 10 µM H89 (PKA inhibitor), 5 nM STP (PKC inhibitor) or 50 µM PD98059 (MEK inhibitor). (b) 2-NBDG uptake in absence/presence of 10 μM Ly294502 (PI3K inhibitor) or 100 nM rapamycin (mTOR inhibitor) (n = 5). RM-one way-ANOVA or ANOVA, with Dunnett’s multiple comparisons test, was used to compare against VIP treatment. Results are expressed as Mean ± S.E.M. *p < 0.05, **p < 0.01, ***p < 0.001.