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. 2021 Mar 31;118(14):e2010210118. doi: 10.1073/pnas.2010210118

Fig. 3.

Fig. 3.

giant responds to the N/C ratio in a kinetic-dependent manner. (A) Heat map of total RNA output for a representative embryo expressing gt>MS2. RNA output is comparable between WT and short-cycle diploid and is lower in haploids. The color bar represents the total cumulative output per nucleus per NC (A.U.). The cartoon shows endogenous gt pattern at NC14, and the dashed box indicates the area under analysis. (Scale bar, 10 µm.) (B) Average gt>MS2 transcriptional trajectory over time for all transcribing nuclei per NC. Data are represented as mean ± SEM of replicate embryos. Gray boxes represent haploid mitoses. (C) Bar chart showing that the number of nuclei transcribing gt>MS2 in a given nuclear cycle is similar between WT (blue), haploid (yellow), and short-cycle diploid (green) embryos. Gray bars represent the total number of nuclei analyzed in each cycle, and colored bars represent the number of active nuclei. Data in B and C are represented as mean ± SEM of six WT, five haploid (ssm), and four short-cycle diploid (grp), biologically replicate gt>MS2 embryos. (D and E) Boxplots showing the rate of transcriptional activation (initial slope) of gt>MS2 for all actively transcribing nuclei for (D) WT versus haploid (ssm) and (E) haploid versus short-cycle diploid (grp) embryos. The initial slope is lower in haploids compared to WT and short-cycle diploids. The dashed line represents the cutoff for outlier values. The numbers of outlier values above the cutoff are given after “+.” Individual data points (up to 200 points) in each nuclear cycle are overlaid on the respective boxplots. **P < 0.005 from Student’s t test. The numbers of nuclei analyzed in C and B, D, and E in each NC are as follows: NC11 [176, 164], NC12 [403, 362], NC13 [830, 730], and NC14 [1,332, 1,077] nuclei from six replicate gt>MS2 WT embryos; NC11 [97, 80], NC12 [311, 270], NC13 [695, 587], NC14 [1,214, 1,013], and NC15 [1,885, 1,513] nuclei from five replicate gt>MS2 haploid embryos; and NC11 [160, 158], NC12 [321, 308], and NC13 [611, 581] nuclei from four replicate gt>MS2 short-cycle diploid embryos.