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. 2018 Sep 13;20(10):1045–1058. doi: 10.1016/j.neo.2018.08.008

Figure 5.

Figure 5

Depletion of Rho suppresses TROY and PDZ-RhoGEF-induced glioma cell migration.

(A) T98G and T98G/TROY-HA cells were transfected with a nontargeting control siRNA (siCTRL) or two independent siRNAs targeting PDZ-RhoGEF (siPRG-1, siPRG-2) for 24 hours and infected with FLAG-tagged PRG lentivirus or GFP lentivirus for additional 24 hours. The cells were serum starved overnight and then seeded on top of a Transwell chamber. The number of migrated cells post 24 hours was quantified using DAPI staining. Data represent the mean values (+/− SEM) (n = 3, *P < .05; **P < .01; ***P < .001). (B) The lysates of T98G used for migration assay in A were immunoblotted with indicated antibodies. (C) The lysates of T98G/TROY-HA used for migration assay in A were immunoblotted with indicated antibodies. (D) T98G and T98G/TROY-HA cells were transfected with a nontargeting control siRNA (siCTRL) or two independent siRNAs targeting PDZ-RhoGEF (siPRG-1, siPRG-2) for 2 days and then serum starved overnight. Cells were lysed, and the Rho activities in lysates were measured using GST-Rhotekin-RBD pull-down assay. Immunoblot is a representation of two independent experiments.