Fig. 6.

Effects of (E)-2-benzylidene-3-(cyclohexylamino)-2,3-dihydro-1H-inden-1-one (BCI), a specific DUSP6 inhibitor, on aldosterone-mediated NCC surface expression and NCC ubiquitination through through ERK1/2 dephosphorylation in mDCT cells. The mDCT cells were pretreated with different concentrations of BCI and then treated with 1 μM aldosterone for 6 h. Cells were then lyzed for Western blot analysis. A: representative blots for the ubiquitinated NCC (Ub-NCC), total NCC, total ERK1/2, and phospho-ERK1/2 were detected by respective antibodies using coimmunoprecipitation and Western blot analysis. B: representative blots for the surface NCC, total ERK1/2, and phospho-ERK1/2 were detected by respective antibodies using cell surface biotinylation and Western blot analysis. C: bar graph representing average band density from 4 independent experiments from A and B. *P < 0.05, **P < 0.01 compared with the control group without aldosterone (Aldo) and BCI treatments.