Comparison between ADAPT and kymograph analysis of blebs. (A) Kymograph of a bleb on a vaccinia-infected HeLa cell expressing GFP and MLC2-RFP, corresponding to the yellow line in Fig. 5 D. (B) Each line represents the membrane displacement (left) and MLC2-RFP intensity (right) derived from one kymograph (representing one bleb; nbleb = 10). (left) The retraction speed was calculated as 1.74 ± 0.08 µm/min by fitting a straight line to the linear portions of the plots representing membrane position over time. The rate of increase in MLC2-RFP intensity was measured as 2.29 × 10−3 ± 1.74 × 10−4 s−1. (C) Analysis of the same cell using ADAPT. Each line represents the mean membrane velocity (left) and MLC2-RFP intensity (right) over time on a single bleb (nbleb = 10). (D) Comparison of mean MLC2-RFP recruitment rates derived from the data in B and C. The rates of increase are 1.95–2.63 × 10−3 and 6.44–6.97 × 10−3 per second for the kymograph- and ADAPT-derived data, respectively. (E) The results of 20 analyses of the same cell as that in B and C using ADAPT, with a different, random set of input parameters for each analysis.
