The diffusion anisotropy is stronger when the posterior domain is photobleached. (A) The quantification of anisotropy index (AI) from the kinetics of fluorescence loss. The anisotropy onset is set as time 0 and fluorescence intensity to 1.0. After fitting the kinetics of loss with one-phase decay, omitting the values after cytokinesis, the time taken for the normalized fluorescence intensity in the A-P region (blue square) to reduce to 0.7 is divided by that in the perpendicular region (red square). The black arrowhead indicates NEBD; the white arrowhead indicates cytokinesis; the orange circle indicates the photobleached area; the asterisk shows the anisotropy onset. Both measurement directions are equidistant from the photobleached area (x). (B) The AIs quantified for wild-type GFP::SP12 (n_anterior = 24; n_posterior = 26) and GFP::KDEL (n_anterior = 10; n_posterior = 11). AIs shown are on log₂ scale. Unpaired t test; *, P < 0.05. (C) The t_1/2 of FRAP in the anterior or the posterior domain of GFP::SP12 embryo. n > 20. Mean ± SD. Unpaired t test, P < 0.05. ns, not significant.