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. 2003 Jan 6;100(2):733–738. doi: 10.1073/pnas.0234057100

Figure 3.

Figure 3

Effect of Tec for Rho activation by Gα12/13-LARG. (A) Activation of SRF activity by Tec. HeLa cells were cotransfected with 0.1 μg of SRE.L-luciferase reporter plasmid with the indicated constructs: 0.01 μg of Gα12QL, 0.01 μg of Gα13QL, 0.1 μg of ΔPDZ-LARG, 0.1 μg of Tec, and 0.1 μg of Tec-KD. SRF activities of cell lysates were measured after 24 h. (B) RhoA activation by Gα12-LARG and Tec in HeLa cells. HeLa cells were transiently transfected with the plasmids encoding ΔPDZ-LARG, Gα12QL, or Tec. GTP-bound RhoA in cell lysates was detected using GST-RBD pull-down assay. The result shown is a representative of three separate experiments with similar results. (C) Direct interaction of Tec with Gα12. COS1 cells were transfected with myc-tagged Tec with or without Gα12QL. Tec was immunoprecipitated from the lysate, and the immunoprecipitates were analyzed by immunoblotting using Gα12 antibody. (D) Requirement of the TH domain of Tec in Gα12/13-LARG signaling pathway. HeLa cells were cotransfected with 0.1 μg of SRE.L-luciferase reporter plasmid with indicated plasmids as described for A or with 0.1 μg of Tec-ΔTH. SRF activities of cell lysates were measured after 24 h.