Figure 8

Involvement of ERK-MAPK pathway in AQP 2 regulation by YHC, GKW and GRA was verified by ERK1/2 specific inhibitor. The mIMCD3 cells were planted in 6 well plate and cultured in RPMI1640 medium supplied with 10% fetal bovine serum. All doses were given during cell logarithmic phase for 24 hours. When the ERK1/2 specific inhibitor U0126 was used, cells were pretreated with U0126 for 1 hour and then for another 24 hours with or without YHC, GKW or GRA. YHC, GKW and GRA all changed AQP 2 expression in mIMCD3 cells, however, when the cells were pretreated with U0126, AQP 2 level was suppressed and these compounds’ activity became not significant compared to U0126 single group. (A,B) Bar chart and (C) blotting image of AQP 2 expression of mIMCD3 cells treated with 12 μM GRA, 8 μM YHC and 25 μM GKW for 24 hours, with or without pretreatment with 10 μM U0126. The blots was cropped properly and the full length blots are presented in Supplementary Fig. 10. *P < 0.05 compared with control group (n = 3).