Extended Data Fig. 6: Additional data for Figs. 7 and 8.
Panels (a–e) are from HEK293T cells. Lanes under the each indicated condition are replicates. a) Distribution of ZAKα across polysome fractions (fractions starting from monosome) related to Fig. 6a. ZAKα abundance in all fractions was normalized to the ZAKα abundance in the first 80S monosome fraction, the first underlined fraction in Fig. 6a. Polysome fractions were numbered from 1 for the first 80S monosome fraction to higher polysome fractions. b) Sucrose gradient polysome profiles without RNase digestion of HEK293T cells under intermediate concentration of emetine (1 µg/ml) treatment. Western blot of ZAKα and ZNF598 distribution in polysome fractions under indicated conditions. ZNF598 in higher polysome fractions are indicated by lines. c) Distribution of ZAKα across polysome fractions (fractions starting from monosome) related to panel b. ZAKα abundance in all fractions was normalized to the ZAKα abundance in the first 80S monosome fraction, the first underlined fraction in panel b. Polysome fractions were numbered from 1 for the first 80S monosome fraction to higher polysome fractions. d) Translation efficiency (TE) of preQ1 treated and control samples, 0Q versus preQ1 from ribo-seq. Highlighted are transcripts whose TE differs by >5-fold between mock and preQ1-treated cells. e) Difference in codon usage between genes with higher TE in preQ1 over 0Q (red transcripts in panel d) and genes with higher TE in 0Q over preQ1 (blue transcripts in panel d). Codon ends with A (red), C (blue), G (orange), U (green) indicated by colors. f) GO term of genes enriched in the ribosome (red mRNA transcripts in panel d) with preQ1. Gene Ontology enrichment analysis was performed using the GeneOntology.org tool with Fisher’s exact test (one-sided). Multiple comparisons were adjusted using the False Discovery Rate (FDR) method. Source numerical data and unprocessed blots are available in.
