Figure 4
Glutamate activates RhoA-ROCK-MLC/MYPT1 pathway. (A) Platelets were stimulated with either thrombin (1 U/ml) for 10 min or 500 µM glutamate (for either 5 or 10 min, in presence or absence of 10 µM Y27632 and 100 µM CNQX) and immunoblotted against p-MYPT1 and p-MLC as indicated. (B,C) Densitometric analysis of immunoblots for p-MYPT1 and p-MLC, respectively, normalized with respect to β-actin. (D) Expression of RhoA-GTP in human platelets stimulated for 10 min with either thrombin or glutamate in presence or absence of Y27632 and CNQX as indicated. Fulllength blots for (A,D) are presented in Supplementary Figs S3 and S4, respectively. (E) Densitometric analysis of RhoA-GTP normalized against total RhoA expression. Bar diagrams represent atleast 5 independent experiments (mean ± SEM). *P < 0.03 as compared to resting platelets; #P < 0.03 as compared to glutamate-stimulated platelets for p-MYPT1. *P < 0.05 as compared to resting platelets; #P < 0.05 as compared to glutamate-stimulated platelets for p-MLC and RhoA-GTP.
