Fig. S3.

Influence of GSK3β inhibition on TRAIL activity in A549 (WT and Dyn1^KO) cells. (A) Immunoblots of two representative single clones. Clone 11 was chosen for additional assays (50, 25, and 12.5 µg of loaded protein per each condition). (B) Levels of surface bound FLAG-tag TRAIL (100 ng/mL) in A549 WT and Dyn1^KO cells after different incubation times at 4 °C. (C) TRAIL effects on cell viability in A549 WT and Dyn1^KO cells, in the presence or absence of the GSK3β inhibitor Chir99021 (10 µM). (D) Effectiveness of Chir99021 after 15-min incubation, as measured by phosphorylated S6K. Cells were preincubated with Chir99021 for 30 min, followed by treatment with several concentrations of TRAIL (1–1,000 ng/mL) and assayed for cell viability. Reduction in cell viability (%) was calculated relative to control wells containing no ligand. All of the results are mean values ± SD (n = 3). Two-tailed Student’s t tests were used to assess statistical significance. ***P < 0.0005.