Fig. 6. Inhibition of RNA Pol I activity is sufficient to induce a fasting-like chromatin architecture in the intestine of fed animals.
a, Top: schematic representation of the degradation kinetics of the core subunits RPC-1 (RNA Pol III), RPB-2 (RNA Pol II) and RPOA-2 (RNA Pol I) in intestine, as quantified in Extended Data Fig. 5a–f. Bottom: single focal planes of representative fed L1 larvae live stained with Hoechst 33342 to monitor DNA, ubiquitously expressing TIR1 and the indicated core subunit endogenously tagged with degron (AID), shown during ethanol exposure as a control, and after 1, 3 and 6 h of 1 mM auxin exposure. Insets: zoom of single nucleus of the intestine. Scale bar, 2.5 μm. b, Quantification of the percentage of the indicated AID-tagged animals (RNA Pol I, Pol II and Pol III) within the indicated categories for 3D chromatin organization in intestine and timepoint of auxin exposure. Data are shown as mean ± s.e.m. of three independent biological replicas. c, Line plots of the averaged single nuclei profile of Hoechst 33342-stained DNA from 72 intestinal nuclei of the indicated fed, degron-tagged animals in presence of ethanol as control or 3 h on 1 mM auxin, from three independent biological replicas, showing the radial fluorescence intensity as a function of the relative distance from the nucleolus centre. The shaded area represents the 95% confidence interval of the mean profile. For RNA Pol I- and Pol II-AID at 3 h of auxin exposure, animals in proportions to their relative abundance within the 3D chromatin organization categories as in b were analysed. d, Line plot as in c but from 70 intestinal cells of fed WT and raga-1GF animals expressing RNA Pol I degron-tagged (RNA Pol I-AID) treated with 1 mM auxin for 5 h. Data are from three independent biological replicas. For c and d, heat maps displaying the single nuclei profiles are provided in Extended Data Figs. 6c,n, respectively. Source numerical data are available in Source data.