Fig. 2.

NOC1 regulates rRNA processing and ribosomal assembling, affecting protein synthesis. (A–C) Representative sucrose density gradient profiles of ribosomes from control larvae (A) or animals over-expressing NOC1 (B) or NOC1-RNAi (C). (D) More detailed view of results shown in A and C highlighting the area of the 40, 60 and 80S ribosomal subunits, noting that the graphs use different scales. (E) Analysis of the percentage of 40, 60 and 80S ribosomal subunits, relative to each genotype, calculated over the total area including the polysome. (F) qRT-PCR showing the fold of induction over control w¹¹¹⁸ of pre-rRNAs analyzed using the ITSs and of mature ribosomal rRNAs; data are expressed relative to actin5C used as control. Results in E and F presented as mean±s.d. for at least three independent experiments. (G) SUnSET western blot analysis of lysates from larvae treated with puromycin for the indicated time. The blot shows the relative changes in protein synthesis using anti-puromycin antibodies in control w¹¹¹⁸ or in larvae ubiquitously expressing NOC1-RNAi under the actin promoter. Actin was used as control loading. (H) Quantification of the change in puromycin incorporation from G and normalized relative to actin (Deliu et al., 2017). Results show mean from two experiments. (I) Ponceau S staining showing total protein levels in G. *P<0.05; **P<0.01; ***P<0.001; ns, not significant (one-way ANOVA with Tukey multi-comparisons test). Images and blots shown are representative of two experiments.