Effects of TDD expression on CCP dynamics. (A and B) Single frame from TIRFM videos (7.5 min/video, see Videos 1 and 2) and corresponding kymographs from region indicated by gray line of adenovirally infected ARPE/HPV cells expressing eGFP-CLCa without (A) and with (B) TDD expression. Arrowheads point to examples of dynamic CCSs subjected to cmeAnalysis. (C) Effect of TDD expression on the growth and stabilization of CCPs, as measured by the rates of initiation of subthreshold CCSs and bona fide CCPs. n = 10 videos; Student’s t test. For box and whisker plots, the boxes encompass the 25th and 75th percentiles, the lines represent the medians, and the whiskers show the minima and maxima data points. (D and E) Mean clathrin fluorescence intensity traces in lifetime cohorts of analyzed clathrin structures (D) and corresponding rate of clathrin recruitment determined by the mean slopes of growth phase (from t = 3 s to t = 8 s; E), indicated by blue arrows in D. Each dot represents the mean slope of a cohort. (F) Maximum fluorescence intensity (I_max) distribution of analyzed CCPs. (G) Lifetime distribution of bona fide CCPs. (H) Median lifetime (LT_50) of analyzed CCPs. Each dot represents a video. Data presented were obtained from a single experiment that is representative of four independent repeats. n = 10 videos for each condition. Number of dynamic tracks analyzed: 124,453 for –TDD and 99,846 for +TDD. Error bars are SD. ***, P ≤ 0.001.