FIGURE 7.

STK17A modulates ROS. A, NT2-STK17Ash2 cells express higher levels of metallothionein genes MT1M, MT1H, and MT1X compared with NT2-PLK control cells as determined by real time PCR. Data points are the average of biological triplicates. Error bars are S.D. *, p < 0.02. B, NT2/D1 cells stably expressing STK17A shRNAs (Sh1 and Sh2) have lower basal and cisplatin-induced ROS levels than control cells (PLK) as measured by the 2′,7′-dichlorodihydrofluorescein diacetate assay. Cells were incubated with or without 1 μm cisplatin for 18 h prior to ROS determination as described under “Experimental Procedures.” Tracings of representative basal ROS determinations in NT2-STK17A-sh1, NT2-STK17A-sh2, and NT2-PLK cells are shown. Bars are the average of two biological replicates, and error bars are the ranges of the two values. The experiment was repeated with similar results. C, 293T cells overexpressing STK17A have higher basal ROS levels. Cells were mock-transfected or transfected with empty vector or an expression plasmid for STK17A, and ROS levels were determined 24 h later. Representative tracings are at left. Bars represent the average of biological triplicate transfections, and error bars are S.D. *, p < 0.005 compared with mock or vector controls. The inset is a Western blot of STK17A overexpression. The experiment was repeated with similar results. M, mock; V, vector.