Fig. 5.

Inhibition of PDBu-induced PKC activation of NKCC1 and ERK1/2 phosphorylation. HCEC were serum starved for 24 h at 80% to 90% confluence. Representative Western blot analysis comparing the effects of exposure to 1 μM PDBu in the presence and absence of either 50 μM bumetanide (BMT) (**p<0.001 PDBu versus treat with BMT). or 10 μM U0126 (**p<0.001 PDBu versus treat with U0126 on phosphorylation levels in both NKCC1 and ERK1/2). on p-NKCC1 and p-ERK1/ 2 detected with anti p-NKCC1 (i.e., R5) and p-ERK1/2 antibodies. Equivalent protein loading was confirmed by reprobing the same blot with anti actin and ERK1/2 antibody, respectively. Data represent the mean ± SEM of three independent experiments.