Extended Data Fig. 2 |. Adaptation to loss of RAB-11.1 through rab-11.2 transcriptional induction.

a, Phenotype enrichment analysis (top 20) of genes differentially regulated (p < 0.05) upon rab-11.1 RNAi. b, Amino acid sequence alignment for RAB-11.1 and RAB-11.2. c,d, Fluorescence micrographs (c) and relative mean fluorescence by flow cytometry (d) of the rab-11.2p::YFP transcriptional reporter in day 1 adults fed ad libitum (control, black) or starved for 24 h (grey). Scale = 200 μm. Mean ± SEM, ***p < 0.0001 by two-tailed unpaired t-test. n = 5 trials (circles). e, Schematic displays the rab-11.2 gene and the respective rab-11.2(syb2999) mutation (731 base pair, bp, deletion). Blue box = exon, blue line = intron. f-i, Representative micrographs (f,h) and relative mean fluorescence by flow cytometry (g,i) of apical transporters, PGP-3::mCherry (f,g) and PEPT-1::DsRed (h,i), in L4 larvae and day 3 adults on EV (black) or rab-11.1 (green) RNAi. Scale = 25 μm. Mean ± SEM, ****p < 0.0001 (g) and **p = 0.0058, ****p < 0.0001 (i) by two-way ANOVA with Sidak’s multiple comparisons test, ns = no significance. n = 3 trials for each timepoint (circles). j, Confocal micrographs of phenotypes representative of TRITC-BSA/FM4–64 absorption efficiency classification categories. Scale = 25 μm. k, Categorization of FM4–64 absorption efficiency following its dietary supplementation in day 2 adult wild type and rab-11.2(syb2999) worms on EV or rab-11.1 RNAi. Shown is the relative ratio of animals which absorbed 0–25% (dark grey), 25–75% (medium grey), or 75–100% (light grey) of ingested FM4–64 into the intestinal epithelium. Mean ± SEM, ****p < 0.0001 by Chi-square test. n = 64 animals per condition over 2 trials. l, Representative micrographs of Oil-Red-O-stained day 5 adult wild type, WT, and rab-11.2(syb2999) worms on EV or rab-11.1 RNAi. n = 3 trials.