Figure 4.
Ribosome production increases when DNA content is limited. (A) Confocal images of NHS-ester stain representing total protein in int2–int3 intestine rings of control and CDK-2gut(−) worms. (B) Quantification of cytoplasmic protein concentration in int2–int4 intestine rings from images as in A. (C) Volcano plot of GO terms for enrichment scores of transcripts in CDK-2gut(−) intestines compared with control. Colored GO terms are related to the nucleosome (lavender squares), translation (orange circles), or transcription (green diamonds). (D) Box plots of residual change in RNA abundance after accounting for expression level (TxPG in control, relationship visualized in Fig. 3 C). Log fold change in TxPG was regressed against expression level using a cubic polynomial. Groups are genes located in the cytoplasm (purple), involved in transcription (RNA Polymerase II subunit or general transcription factor; green) or ribosome subunits (orange). Kruskal–Wallis test P = 1.0 × 10−13; P values shown are from post-hoc Dunn test with Benjamini-Hochberg multiple hypothesis testing correction. (E) Quantification of change in TxPG of the 18S or 5S rRNAs in control and CDK-2gut(−) intestines. Each connected pair of points represents one paired biological replicate. The mean TxPG of control is scaled to 1. P values are from paired t test with Benjamini-Hochberg multiple hypothesis testing correction. (F) Representative western blot showing total protein and anti-RPS-5 signal in dissected control and CDK-2gut(−) intestines. (G) Quantification of anti-RPS-5 signal normalized to total protein as in F, with control ratios for each blot scaled to 1. Each dot represents one experiment (N = 3 blots, n = 50–80 worms per genotype per blot). P value is of one sample t test against null hypothesis that μ = 1. (H) Confocal images of RPS-6::mCherry in int2–int4 intestine rings of control and CDK-2gut(−) worms. (I) Quantification of cytoplasmic RPS-6 concentration in int2–int4 intestine rings from images as in H. In B and I, each dot represents one worm, bars and error bars represent means and 95% confidence intervals, and P values are from Mann-Whitney U tests. Source data are available for this figure: SourceData F4.

Ribosome production increases when DNA content is limited. (A) Confocal images of NHS-ester stain representing total protein in int2–int3 intestine rings of control and CDK-2gut(−) worms. (B) Quantification of cytoplasmic protein concentration in int2–int4 intestine rings from images as in A. (C) Volcano plot of GO terms for enrichment scores of transcripts in CDK-2gut(−) intestines compared with control. Colored GO terms are related to the nucleosome (lavender squares), translation (orange circles), or transcription (green diamonds). (D) Box plots of residual change in RNA abundance after accounting for expression level (TxPG in control, relationship visualized in Fig. 3 C). Log fold change in TxPG was regressed against expression level using a cubic polynomial. Groups are genes located in the cytoplasm (purple), involved in transcription (RNA Polymerase II subunit or general transcription factor; green) or ribosome subunits (orange). Kruskal–Wallis test P = 1.0 × 10−13; P values shown are from post-hoc Dunn test with Benjamini-Hochberg multiple hypothesis testing correction. (E) Quantification of change in TxPG of the 18S or 5S rRNAs in control and CDK-2gut(−) intestines. Each connected pair of points represents one paired biological replicate. The mean TxPG of control is scaled to 1. P values are from paired t test with Benjamini-Hochberg multiple hypothesis testing correction. (F) Representative western blot showing total protein and anti-RPS-5 signal in dissected control and CDK-2gut(−) intestines. (G) Quantification of anti-RPS-5 signal normalized to total protein as in F, with control ratios for each blot scaled to 1. Each dot represents one experiment (N = 3 blots, n = 50–80 worms per genotype per blot). P value is of one sample t test against null hypothesis that μ = 1. (H) Confocal images of RPS-6::mCherry in int2–int4 intestine rings of control and CDK-2gut(−) worms. (I) Quantification of cytoplasmic RPS-6 concentration in int2–int4 intestine rings from images as in H. In B and I, each dot represents one worm, bars and error bars represent means and 95% confidence intervals, and P values are from Mann-Whitney U tests. Source data are available for this figure: SourceData F4.

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