DualTag-based imaging allows monitoring the dynamic transcriptional changes of icl-1 and acdh-1 in the intestinal cells. (A) Representative images to show the level of icl-1 nascent mRNAs and ICL-1::MCHERRY proteins in the intestinal cells. L4 stage worms were fasted from 0 to 8 h. Arrowheads indicate RNA spots. z-stacks were captured and proper single-focal-plane images were shown. Scale bars: 5 μm. (B) Quantification of the percentage of the nucleus with RNA spots of icl-1 in A. Each dot represents a single worm. n represents the number of animals. (C) Quantification of the intensity of MCP::GFP-labeled RNA spots per nucleus indicating total nascent transcripts of icl-1 under the conditions showed in A. Each dot represents a single intestinal cell. n represents the number of intestinal cells. (D) Quantification of protein expression level by measuring the fluorescence intensity for each condition in A. n represents the number of animals. (E) Representative images to show simultaneous visualization of nascent RNAs and proteins of acdh-1 in the intestine during fasting. L4 stage worm were selected and then fasted for 0, 2, 4, 6, and 8 h before imaging. Arrowheads: nascent RNAs. Proper single-focal-plane images were shown. Scale bars: 20 μm. (F) % of nucleus with RNA spots were quantified to report the transcription activity of each worm in E. Each dot represents a single worm. n represents the number of animals. (G) Quantifications of transcriptional output per cell were performed as those in C. Each dot represents a single cell. n represents the number of intestinal cells. (H) Quantification of ACDH-1::MCHERRY protein level, similar to D. n represents the number of animals. Data = mean ± SEM. ns: not significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way ANOVA with the Tukey correction).