Fig. 5
Zelda global kinetic properties. a Living GFP-zld/ + ;His2Av-mRFP/ + embryo imaged by confocal microscopy from interphase of nc13 to early interphase of nc14. Successive representative maximum intensity projected Z-stack images are shown at the indicated timings (in minutes) (see also Supplementary Movie 9). Scale bars represent 20 μm. b Average intensity profiles for histones (red), nucleoplasmic Zelda (green) and cytoplasmic Zelda (blue) measured from a nc13 embryo transitioning into nc14. An automatic tracking of fluorescence from a minimum of 114 nuclei and 10 cytoplasmic ROI generated these profiles. Error bars represent SD. c FRAP mean curve (black) and the mean of all the fits (red curve) using reaction-diffusion models determined at the bleached spot for 25 nuclei from nc14 developing GFP-zld embryos. Error bars represent SD from different nuclei (light blue bars). d Example of a time trace obtained by FCS from a GFP-zld nc14 embryo showing no bleaching. e Example of autocorrelation function (black dots) related to d (red curve represents fitting using a reaction-diffusion model). f–g Kinetic parameters for Zelda and Ash1 extracted after fitting FCS data with a reaction-diffusion model. f Box plot representing estimated diffusion coefficients (Df). Numbers above each plot represent the mean. Centered lines represent the median and whiskers represent min and max. g Box plot representing estimated residence times (1/koff). Numbers above each plot represent the mean. Centered lines represent the median and whiskers represent min and max
