The graphs are side-by-side and separate, each representing different data sets. The horizontal axis represents time in minutes, and the vertical axis represents relative intensity in arbitrary units (AU). Panel A: A line graph showing normalized relative intensity curves of H3.2-Dendra2 across nuclear cycles 11 to 13 in wild-type embryos. The horizontal axis represents time in minutes, and the vertical axis represents normalized H3.2-Dendra2 intensity, with values normalized to the maximum intensity in nuclear cycle 11. Panel B: A line graph showing normalized relative intensity curves of H3.2-Dendra2 across nuclear cycles 11 to 13 in NASP-deficient embryos, plotted similarly with intensity normalized to nuclear cycle 11. Panel C: A scatter plot showing normalized nuclear import rates of H3.2-Dendra2 across nuclear cycles, comparing wild-type embryos in blue and NASP-deficient embryos in red, with a simulated condition shown in black. Data are presented as mean with standard deviation, and statistical significance is indicated by asterisks denoting different P-value thresholds. Panel D: A line graph showing normalized relative intensity curves of H3.3-Dendra2 across nuclear cycles 11 to 13 in wild-type embryos, with values normalized to the maximum intensity in nuclear cycle 11. Panel E: A line graph showing normalized relative intensity curves of H3.3-Dendra2 across nuclear cycles 11 to 13 in NASP-deficient embryos, plotted similarly. Panel F: A scatter plot showing normalized nuclear import rates of H3.3-Dendra2 across nuclear cycles, comparing wild-type and NASP-deficient embryos, with data presented as mean with standard deviation and statistical significance indicated. Panel G: A line graph showing simulated normalized relative intensity curves for histone H3 nuclear import at reduced initial concentration, normalized to nuclear cycle 11 maximum intensity. Panel H: A line graph showing slopes used to estimate nuclear import rates from the simulation data over time. Panel I: A scatter plot showing cell cycle duration across nuclear cycles 11 to 13 in wild-type and NASP-deficient embryos, including a simulated condition, with values presented as mean with standard deviation and statistical comparisons indicated. Panel J: A line graph showing H3.2-Dendra2 total nuclear intensity over time, comparing wild-type and NASP-deficient embryos, with shaded regions indicating specific experimental phases. Panel K: A line graph showing H3.3-Dendra2 total nuclear intensity over time, comparing wild-type and NASP-deficient embryos, with similar temporal segmentation indicated by shaded regions.
Normalized nuclear import rates for H3.2 and H3.3 Dendra. (A) Normalized relative intensity curves of H3.2-Dendra2 for nuclear cycles (NC) 11–13 in WT embryos. Values normalized to maximum H3.2-Dendra2 intensity in NC11. (B) Normalized relative intensity curves of H3.2-Dendra2 for nuclear cycles (NC) 11–13 in NASP-deficient embryos. Values normalized to maximum H3.2-Dendra2 intensity in NC11. (C) Normalized nuclear import rates of H3.2-Dendra2 in WT (blue) and NASP-deficient (red) embryos. Simulated import rate for H3.2 at 50% WT levels (black). N = 6 biological replicates. Data represent the mean ± standard deviation (SD). Statistical difference between groups were analyzed by employing a two-way ANOVA Tukey’s multiple comparison test between the WT and NASP data points. (D) Normalized relative intensity curves of H3.3-Dendra2 for nuclear cycles (NC) 11–13 in WT embryos. Values normalized to maximum H3.2-Dendra2 intensity in NC11. (E) Normalized relative intensity curves of H3.3-Dendra2 for nuclear cycles (NC) 11–13 in NASP-deficient embryos. Values normalized to maximum H3.2-Dendra2 intensity in NC11. (F) Normalized nuclear import rates of H3.3-Dendra2 in WT (blue) and NASP-deficient (red) embryos. N = 6 biological replicates. Data represent the mean ± standard deviation (SD). Statistical difference between groups were analyzed by employing a two-way ANOVA Tukey’s multiple comparison test between the WT and NASP data points. (G) Normalized relative intensity curves for simulation of H3 nuclear import at 50% initial concentration. Values normalized to maximum H3.2-Dendra2 intensity in NC11. (H) Slope for nuclear import rate calculation for simulation of H3 nuclear import at 50% initial concentration. (I) Cell cycle duration for NC11-13 in WT (blue) and NASP-deficient (red) embryos. Simulated cell cycle duration for NC13 at 50% WT H3.2 levels (black). N = 6 biological replicates. Data represent the mean ± standard deviation (SD). Statistical difference between groups were analyzed by employing a two-way ANOVA Tukey’s multiple comparison test between the WT and NASP data points. In the figure panels, ns represents nonsignificant difference, while *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001. (J) Un-normalized nuclear export measurement for H3.2-Dendra2 in WT and NASP-deficient embryos in NC12. Time point 0 represents Nuclear Envelope Breakdown (NEB). N = 3 biological replicates. Data represents mean ± standard deviation (SD). (K) Un-normalized nuclear export measurement for H3.3-Dendra2 in WT and NASP-deficient embryos in NC12. Time point 0 represnts Nuclear Envelope Breakdown (NEB). N = 3 biological replicates. Data represents the mean ± standard deviation (SD).