GTSE1 modulates cell migration in an EB1-dependent manner.

<p>(A) Transwell migration assay and western blot in U2OS cells, and U2OS cells stably expressing RNAi-resistant wild-type GTSE1-GFP (GTSE1^WT₂₀₄), or RNAi-resistant GTSE1-GFP mutated at the SKIP motifs (GTSE1^Sk₂₀₂). Cells were transfected with control (CON) or GTSE1 siRNA for 36 h and seeded on transwell membranes. Histograms show the mean number of cells/area that migrated through the transwell after 16 h (10 replicates/experiment). Error bars represent the standard error of the mean from three independent experiments. Western blots were performed on cells after the same treatment, and blotted with anti-GTSE1 and anti-actin. (B) Transwell migration assay and western blot in Wi38 cells. Cells were transfected and analysed as in (a). (C) Transwell migration assay and western blot in HCT116 wild type, HCT116 p53−/−, and HCT116 p21−/− cell lines. Cells were transfected and analyzed as in (a). Western blots were performed on cells after the same treatment, and blotted with anti-GTSE1, anti-p21, anti-p53, and anti-actin. (D) Transwell migration assay and western blot in H1299 cells containing inducible constructs for expression of wild type GTSE1 (GTSE1 WT) or SKIP-domain mutated GTSE1 (GTSE1 Sk). Cells were untreated (−) or treated with Ponasterone A (PonA) (+) for 24 h to induce GTSE1 expression, then trypsinized and seeded on transwell membranes. Histograms show the mean number of cells/area that migrated through the transwell after 16 h (10 replicates/experiment). Western blots were performed on cells after the same treatment, and blotted with anti-GTSE1 and anti-actin.