Fig. 1: Feeding stimulates protein synthesis and triggers UPRER activation.
a Schematic overview of the methods used to measure protein synthesis and gene expression in response to feeding. b Western blot analysis showing puromycin incorporation in animals with or without food. The SUnSET assay was used to monitor ongoing protein translation, detected by puromycin incorporation into newly synthesized proteins. A significant increase in puromycin levels was observed in L1-stage animals fed with OP50 for 10 h (*p < 0.05, two-sided t-test). Error bars represent the mean ± SD of three independent biological replicates. c Gene Ontology enrichment analysis of food-induced genes in L1-stage animals. Animals were grown on NGM plates for 4 h with or without OP50 feeding. The raw p-values were adjusted to obtain the padj values. d Heatmap displaying the clustering of translation-related gene expression in animals with or without food. Colors indicate the relative intensity of gene mRNA levels (Red: high; Blue: low) in the presence or absence of OP50. e Quantification of hsp-4p::gfp fluorescence in L1 animals after varying durations of feeding (L1 + 0.5 h, L1 + 1.5 h, L1 + 3 h, L1 + 6 h, L1 + 8 h, L1 + 24 h). Mean ± SD; ns denotes p > 0.05, ****p < 0.0001 via two-sided t-test. f Fluorescent micrographs and quantification of GFP in L1 transgenic animals expressing hsp-4p::gfp after 24 h of feeding. Scale bar, 20 μm. Mean ± SD; ****p < 0.0001 via two-sided t-test. g Schematic illustration depicting the relationship between food digestion and ER homeostasis. Food digestion enhances protein synthesis, which may activate the UPRER to mitigate ER overload and maintain ER protein homeostasis. However, it remains unclear whether disruptions in ER protein homeostasis could feedback into the food response system, potentially leading to the inhibition of food digestion to reduce ER stress. For all panels, n = number of animals which were scored. Representative data shown is one of three biological replicates. Source data are provided as a Source Data file. See also Supplementary Figs. 1 and 2 and Supplementary Data 1.
