Fig. 1: DFR charge controls the distribution of TF residence times.
a Net charge in DBD (red) and non-DBD (black) of Sox TFs. b Left, Pearson correlation between the mitotic bound fraction (MBF)40 and the net charge in non-DBD of Sox TFs in a; Right, representative images40 of H2B-mCherry, Sox2-YPet (top) and Sox17-YPet (bottom) localization in metaphase. Scale bar: 10 μm. c Charge distributions for Sox2 (top) and Sox17 (bottom), calculated as sliding-window averages of residue charges over five-amino-acid segments (see the “Methods” section). d Scheme representation of the Sox TF library. e Top, representative images of in vivo SMT. Nuclei (left, red) and sparsely labeled Sox TFs (right, yellow) are visualized in the SiR647 and JF549 emission channels, respectively. Scale bar: 15 μm; Bottom, scheme of in vivo single-molecule imaging using HILO microscopy (left) and a labeling strategy for Sox TFs using the Halo-JF549 dye (right). f Representative images of JF549-labeled Halo-Sox2 molecules, acquired at different time intervals in time-lapse movies. Scale bar: 2 μm. The kymographs below each image correspond to the molecules in the highlighted squares. g Reverse cumulative density function (1-CDF) of Sox TF bound times. Curve shadings: standard deviation (SD) calculated from 100 resampling iterations, each using a randomly selected subset (80%) of the full dataset selected randomly (N = 281, 169, 223, 91, 164, 159 cells). h Residence time distributions obtained from GRID analysis of 100 resampling iterations (randomly selected 80% subset of the full dataset shown in g), assembled into histograms (see the “Methods” section). The GRID amplitude indicates the bound fraction per unit of time for the corresponding residence times.
