DualTag monitors the dynamic transcriptional changes of metabolic genes in response to fasting. (A) Representative images illustrating the levels of icl-1 nascent mRNAs and ICL-1::MCHERRY proteins in the skin cells without or with fasting treatment. L4 stage animals were collected and analyzed. (B) Quantification of % of nuclei with RNA spots, number of transcriptional active alleles per nucleus, the intensity of single RNA spot, and protein intensity of icl-1 under different conditions as shown in A. When quantifying the % of nuclei with RNA spots and protein intensity, n represents the number of animals. In the analysis of the intensity of single RNA spot, n represents the number of alleles. n represents the number of nuclei for quantifying number of transcriptional active alleles per nucleus. The exact numbers analyzed were indicated above each bar. (C and D) Similar to the analysis in A and B, but the fasting duration for adh-1 ranged from 0 to 12 h. (E and F) Similar to A and B except that acdh-1 was analyzed. L4 stage animals were imaged after fasting for 0, 2, 4, 6, or 8 h respectively. (G and H) Similar to A and B except that hach-1 was analyzed. L4 stage worms were well-fed or fasted 2–8 h. All images in Fig. 5 were captured by z-stacks and proper single-focal-plane images were shown. Arrowheads: nascent RNA. Scale bars: 5 μm. All data are presented as mean ± SEM. ns: not significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 (one-way ANOVA with the Tukey correction).