Applying Automated Tracking of Microtubules by plusTipTracker

Abstract Automated microtubule tracking programs have potential as a powerful tool for analyzing a variety of experimental conditions. Here, we apply one such program, plusTipTracker, developed by the Danuser lab, to investigate the effects of stathmin and p27 depletion on microtubule dynamics. We also apply plusTipTracker to analyze the relationship between p53 and taxol on microtubule dynamics. By developing a standardized procedure, we were able to streamline the detection and analysis of these conditions much faster than is possible by hand. We found that both p27 and stathmin depletion alter microtubule dynamics. This is proposed to be a result of p27 binding to stathmin and competeing for stathmin's tubulin-sequestering abilities in vivo. p53 was found to repress microtubule dynamics in interphase HeLa cells. Taxol was also found to greatly repress microtubule dynamics, and this repression is augmented by the presence of p53. Based on these observations and results, we conclude plusTipTracker is a powerful method for the analysis of microtubules under our conditions.