Fig. 6.

ROCK1 promoted TMEM16A channel activity by phosphorylating moesin at T558. A. Time course of Ca²⁺ (1 μM)-activated Cl⁻ currents in T47D cells treated with the ROCK inhibitor Y-27632 (10 μM). Voltage ramps 750 ms in duration were induced at 10-s intervals. The arrow indicates treatment with Y-27632. The currents were normalized to the initial values of current prior to Y-27632 treatment. B. Representative TMEM16A currents before (top) and after (bottom) Y-27632 treatment. The cells were voltage-clamped with a 750-ms voltage step from –100 mV to +100 mV in 20 mV increments. C. Mean current densities at +100 mV in T47D cells before and after Y-27632 treatment. n = 5 cells. *p < 0.05 vs before treatment. D. Expression of p-ROCK1, ROCK1, p-moesin (T558), and moesin in T47D cells overexpressing RhoA-V14 and RhoA-19 N mutants. E. Representative TMEM16A currents in T47D cells transfected with plasmids containing RhoA-V14 and RhoA-19 N mutants. F. Mean current densities at +100 mV in T47D cells treated with RhoA-V14 and RhoA-19 N mutants. n = 5–6; *p < 0.05. G. I. Representative TMEM16A currents in response to activation by 1 μM Ca²⁺ in T47D cells (G) transfected with plasmids containing T558D-moesin or T558A-moesin mutants or in HEK293 cells (I) cotransfected with plasmids containing TMEM16A and T558D-moesin or T558A-moesin mutants. H, J. Mean current densities at +100 mV in T47D (H) and HEK293 cells (J). n = 4–5; *p < 0.05.