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{"CAPTION 2.png": "'Figure 1: **Cell processing blocks that and assemble skeletons in multiple cell operators.** (3) (Cutter and temperature) and temperature levels of a 3DCT cell are shown. **Fig. 2:**_Cell processing blocks that and assemble skeletons in multiple cell operators.** (4) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 3:**_Cell processing blocks that and assemble skeletons in multiple cell operators.** (5) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 4:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (6) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 5:**_Cell processing blocks that and assemble skeletons in multiple cell operators.** (7) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 6:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (8) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 7:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (9) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 8:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (10) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 9:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (11) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 10:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (12) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 11:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (13) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 12:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (14) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 13:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (15) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 14:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (16) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 15:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (17) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 16:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (18) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 17:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 18:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (10) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 19:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (11) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 10:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (12) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 11:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (13) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 12:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (14) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 13:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (15) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 14:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (16) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 15:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (17) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 16:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (18) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 17:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 18:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 19:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (11) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 11:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (12) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 12:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (13) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 13:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (14) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 14:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (15) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 16:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (16) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 17:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (17) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 18:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (18) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 19:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 10:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 11:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 12:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 13:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 14:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 15:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 16:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and temperature levels of a 3DCT cell are shown). **Fig. 17:**_Cell processing blocks and assemble skeletons in multiple cell operators.** (19) (Cutter and\\n\\n'", "CAPTION 2F.png": "'\\n\\n## References\\n\\nFig. 2: **Cell squeezing induces fast and reversible alterations in clathrin coat dynamics.** (A) Cartoon and representative frames of a BSC1 cell are shown at different stages of squeezing (Movie 2). (B) Kymograph showing the temporal evolution of the clathrin traces detected at the ventral surface of the cell shown in A. Dashed lines mark the squeezing steps. (C) For the cell in A, normalized distributions of clathrin growth rates are plotted for different squeezing levels (Fig. S1). The standard deviation of the distribution reduces as the tension increases. (D) For the same cell, the time variation of the ventral surface area (upper) and the standard deviation of the clathrin growth rates (lower). The stepwise changes in these parameters are due to discrete levels of squeezing. (E) The response of the same cell to squeezing is shown as the mean clathrin lifetime (upper) and initiation and conclusion densities (lower). Dashed lines indicate changes in squeezing (\\\\(n_{\\\\text{vacuum}}\\\\)=8217). (F) Standard deviation of clathrin growth rates (upper), mean lifetime (middle), and initiation and conclusion densities (lower) from a cell that undergoes increased stepwise squeezing (orange dashed lines) and relaxation (blue dashed lines) (Movie 3) (\\\\(n_{\\\\text{vacuum}}\\\\)=8255).\\n\\n'", "CAPTION FIG1.png": "'\\nFigure 1: **Aspiration of the plasma membrane slows down clathrin coat dynamics.** (A) Kymograph showing the clathrin activity at the ventral surface of a BSC1 cell expressing AP2-eGFP. Clathrin coat traces elongate gradually upon microsuspension (dashed line; Movie 1). Blue and red arrowheads mark the initiation and conclusion of a clathrin-coated structure, respectively. _A)_ its its lifetime. (B) Clathrin coat lifetime distributions are shown for nine BSC1 cells imaged before and during microsuspension (\\\\(n_{\\\\text{max}}\\\\)=40,943). (C) For the same nine cells, the standard deviation of the clathrin growth rate distributions are shown in koptics. Lines connect the standard deviation values obtained from the same cell before and during aspiration. The narrower growth rate distributions indicate slower clathrin coat dynamics. (D) Box plots are the initiation and conclusion densities of clathrin-coated structures before and during aspiration. In the koptics, the box represents the 25\u201375th percentiles, and the median is indicated. The whiskers show the 10\u201390th percentiles. _P_-values were obtained with a two-tailed _I_-test.\\n\\n'", "CAPTION FIG3.png": "'Figure 3: Hypetemic swelling inhibits clathrin coat dynamics temporarily. (A) Change in the volume (normalized to the radial value) is placed for three ESC1 cells during hypotonic swelling (i.e. osmachock). (B) Mean clathrin coat lifetime (upper), and initiation and conclusion densities (lower) are plotted against time for a ESC1 cell treated with hypotonic shock (dashed line). (C) Clathion lifetime distributions are assembled pre- and post-osmachock for 12 gene-edited SUM159 cells xenrazing AP2-EGFP (\\\\(n_{\\\\mathrm{tracea}}\\\\)=\\\\(34,113\\\\)). (DE) For the same cells, the standard deviation of clathrin growth rates [0] and initiation and conclusion densities of clathrin-coated structures pre- and post-osmotic shock(E) are shown in boxplots. Lines centered the standard deviation values obtained from the same cell pre- and post-osmachock. In the boxplots, the box represents the 25-75th percentiles, and the median is indicated. The whiskers show the 10-90th percentiles. Values were obtained with a two-tailed \\\\(t\\\\)-test.\\n\\n'", "CAPTION FIG4.png": "'\\nFigure 4: **Acth dynamics mediate the inward movement of clathrin coats prior to disassembly.** (A) Means-s.e.m. values for of normalized AP2 intensity traces (determined for a SUM159 cell before and after hypotonic swelling; \\\\(n_{\\\\text{max}}\\\\)=3728). The traces are aligned at the end point before averaging. (B) Mean-s.e.m. z. displacements are shown for the two trace groups in A. (C) Top, growth rate distributions are assembled for eight SUM159 cells before and after hypotonic swelling (\\\\(n_{\\\\text{max}}\\\\)=30,409). Different growth phases (ff, fast formation; sf, slow formation; p, plateau; sd, slow dissolution; fd, fast dissolution) were determined by quantifying the change in the clathrin coat signal over 25-s long time windows (Ferguson et al., 2016). Bottom, for the same cells, bar plots show the mean-s.e.m. of the z. velocities of the trace fragments (12 s long) that are used to generate the growth rate distributions above. Trace fragments that have the highest z velocity are found in the fast dissolution (1fd) phase. (D) Top, representative intensity traces of AP2 (green) and LifeAct (red) fluorescence during the formation of a clathrin-coated vesicle at the ventral surface of a BSC1 cell. Bottom, the relative LifeAct intensity (mean-s.e.m.) colocalizing with clathrin coats is shown for different growth phases (\\\\(n_{\\\\text{total}}\\\\)=4, \\\\(n_{\\\\text{max}}\\\\)=28,796). Note that the growth phases are determined by using the master (AP2=EGFP) signal. (E) Bar plots show the z velocities (mean-s.e.m.) corresponding to different growth phases for AP2 traces obtained from BSC1 cells in the absence and presence of jasplakinolide (Jase) (Control, \\\\(n_{\\\\text{total}}\\\\)=7, \\\\(n_{\\\\text{max}}\\\\)=20,204; Jase, \\\\(n_{\\\\text{max}}\\\\)=6, \\\\(n_{\\\\text{max}}\\\\)=25,972). *P-O0.0001; *P-O0.001 (two-tailed Hest).\\n\\n'"} |
