Figure 7

Pharmacological inhibition of Rho-dependent kinase (ROCK) prevents OI-V-induced disruption of apical junctions. HPAF-II cells were treated for 5 h with either OI-V alone (1 μM) or in a combination with two pharmacological ROCK inhibitors, Y-27632 (20 μM) and H-1152 (10 μM). The effects of the PKC activator on barrier properties were determined by TEER measurement (A), whereas the integrity of AJ and TJ was analyzed by immunofluorescence labeling for E-cadherin and occludin respectively (B). Both ROCK inhibitors significantly attenuate OI-V-induced disruption of paracellular barrier and disassembly of AJ and TJ. Bar, 20 μm. (C) Immunoblotting analysis shows that inhibition of ROCK using H-1152 (10 μM) abolishes both mono- and di-phosphorylation of RMLC induced by 3 h exposure to OI-V (1 μM), while not affecting the levels of total RMLC.