mRNA abundance is partially limited by DNA content in the intestine. (A) Confocal images of FISH against poly(A) tails in int2–int3 intestine rings of control and CDK-2^gut(−) worms. (B) Quantification of cytoplasmic mRNA concentration in int2–int4 intestine rings from images as in A. (C) Quantification of the ratio of the nucleoplasmic mRNA to the cytoplasmic mRNA from images as in A. In B and C, each dot represents one worm. (D) Live confocal images of intestinal nuclei of day 1 adult worms expressing endogenously-tagged Pol II (AMA-1::GFP). (E–G) Quantification of the total volume (E), total amount (F, total fluorescence intensity), and concentration (G, mean fluorescence intensity) of AMA-1 in int2–int4 cells. Volume and total intensity measurements of individual nuclei were summed in binucleate cells. Each dot represents one cell. In B, C, and E–G, bars and error bars represent means and 95% confidence intervals; P values are from Mann-Whitney U tests after Benjamini-Hochberg multiple hypothesis testing correction. (H) Cartoon depicting normalized RNA-seq experiment: dissected intestines from two species were mixed, and C. elegans gene expression was normalized to the spike-in species, P. pacificus. (I) Distribution of transcripts per genome (TxPG) across all genes in control and CDK-2^gut(−) intestines. P value is from t test after log transformation of TxPG values. (J) Distribution of changes in TxPG of all genes. The solid purple line represents the median change (Δ) in expression, and dashed black line indicates the level of expression change that would be necessary to fully restore normal mRNA concentration (1/ploidy reduction).