FTI-277 treatment of HeLa cells increases focal adhesions. A: The images show the localization of vinculin in FTI277-treated and vehicle-treated samples together with the software mask to identify total vinculin (red) and vinculin at the focal adhesions (blue). HeLa cells were treated as indicated in the text and stained for immunofluorescence analysis with α-vinculin primary and appropriate secondary fluorescently labeled antibody, as indicated in Methods. To calculate the number of focal adhesions in each experiment at least 86 cells were counted per sample. The software mask corresponding to the vinculin signal is highlighted (panel A, Mask, blue circled spots). The number of focal adhesions present in each sample was counted and normalized against the total number of cells counted. The number of focal adhesions in vehicle samples was arbitrarily considered 100%. The yellow scale bar represents 10 μM. B and C. The graphs show the mean ± SD of at least 2 independent experiments, each run in triplicate (three wells per condition) at the indicated time points.