Every Little Thing You Don't Know About PTEN Might Probably Surprise You
Movies were made with a time lag of 3?s and 300?ms exposure, using 488?nm laser light. For the analysis of MT dynamics, DIC data were collected from three independent experiments for every condition. The plus-ends of the axonemes were identified by the presence of longer and faster growing MTs (Walker et?al., 1988). Only plus-ends were analyzed. To determine the percentage of buckling/straight MTs in contact with the barrier (Figure?1), we visually counted the number of buckling versus straight MTs in the movies. To determine the time MTs spend ��searching�� along the barrier, we recorded the total time MT ends were (visually) observed to slip along the barrier and divided this by the total number of observed capture events. We found average search times of 220?s for 100% dynein (10 events), 670?s for 10% dynein (18 events), 890?s for 1% dynein (22 events), and 2705?s in the absence of dynein (4 events), indicating selleck screening library an increase in search time with decreasing dynein density (although the values for 1% and 10% are not significantly different; p-value 0.14, see definition below). For a subset of MTs, we compared this buy TSA HDAC visual analysis to a more quantitative analysis as described in Figure?S1. Here, we quantified individual MT behavior over time to determine if they were buckled or straight, and to determine whether their ends were sliding or stationary along the barrier. We did not observe any significant differences in the results. Catastrophe, barrier release, and rescue frequencies (Figure?1, Figure?2, and Figure?S2) were determined by dividing the total number of observed catastrophe, release, or rescue events by the total time that the MTs were (visually) observed to grow (either free or gliding/buckling against a barrier), remain straight in stationary contact with the barrier, or shrink (only for free MTs). The PTEN error is the statistical error given by the frequency itself divided by the square root of the number of observed events. For n?
