Panel A: Time-lapse fluorescence images showing nuclear dynamics of H3.2-Dendra2 (left) and NASP-Dendra2 (right) across successive time points from one mitotic stage to the next, illustrating differences in accumulation patterns within nuclei. Panel B: A line graph showing relative intensity of NASP-Dendra2 over time across nuclear cycles 11 to 13. The horizontal axis represents time in minutes, and the vertical axis represents normalized relative intensity. An adjacent scatter plot shows import rates across nuclear cycles, with the horizontal axis representing nuclear cycle (11–13) and the vertical axis representing import rate, with statistical significance indicated. Panel C: A line graph showing relative intensity of H3.2-Dendra2 over time across nuclear cycles 11 to 13. The horizontal axis represents time in minutes, and the vertical axis represents normalized relative intensity. An adjacent scatter plot shows import rates across nuclear cycles, with the horizontal axis representing nuclear cycle (11–13) and the vertical axis representing import rate, with statistical significance indicated. Panel D: A scatter plot showing the percent decrease in nuclear import rate relative to nuclear cycle 11 for both H3.2-Dendra2 and NASP-Dendra2. The horizontal axis represents nuclear cycles (11–13) grouped by protein type, and the vertical axis represents percent decrease in import rate, with statistical comparisons indicated between groups.
H3 and NASP have different nuclear import kinetics. (A) Time-lapse of H3.2-Dendra2 (left) and NASP-Dendra2 (right) from metaphase 12 to metaphase 13. Scale bar represents 25 µm. (B) Relative intensity curves of NASP-Dendra2 for nuclear cycles NC 11–13 in WT embryos. N = 7 biological replicates. Data represent the mean ± standard deviation (SD). Statistical difference between the groups were analyzed by employing a one-way ANOVA Tukey’s multiple comparison test. (C) Relative intensity curves of H3.2-Dendra2 for nuclear cycles NC 11–13 in WT embryos (same as in Fig. S1 A). N = 6 biological replicates. Data represent the mean ± standard deviation (SD). Statistical difference between the groups was analyzed by employing a one-way ANOVA Tukey’s multiple comparison test. (D) The percent decrease in the nuclear import rate of H3.2-Dendra2 and NASP-Dendra2 for NC 11–13. N > 6 biological replicates. Data represent the mean ± standard deviation (SD). Statistical difference between the groups was analyzed by employing a two-way ANOVA with Geisser–Greenhouse correction and Fisher’s least significant difference (LSD) to identify pairwise differences. In all figure panels, ns represents nonsignificant difference, while **P < 0.01, ***P < 0.001 and ****P < 0.0001.