Extended Data Fig. 5. The microbiota of hodor mutants.

a, Increased bacterial loads (CFU/larvae) in hodor mutants when compared to control larvae. Bacterial loads were assessed in third-instar larvae raised on a high-yeast diet. b-c, Developmental rate of control and hodor mutant larvae in germ-free conditions, or following re-colonisation with Acetobacter pomorum or Lactobacillus plantarum in either high (b) or low-yeast (c) conditions. hodor mutants remain developmentally delayed in germ-free conditions, particularly when reared on a low-yeast diet. Mono-association partially rescues the developmental delay of all larvae in low-yeast conditions, but the difference in developmental rate between control and hodor mutant larvae persists. d, Representative images of FluoZin-3AM stainings (a zinc dye) in the copper cell region of larvae reared in germ-free conditions or bi-associated with Acetobacter pomorum and Lactobacillus plantarum. More zinc is apparent in the copper cell region of high yeast-fed larvae relative to low yeast-fed larvae, but this is unaffected by the presence of microbiota. e, Quantifications of zinc staining in copper cell region. See Supplementary information for sample sizes and full genotypes. Scale bars: d, 30μm. For comparisons involving two groups, a non-parametric Mann Whitney U test was used. Where more than two groups were compared, an ordinary one-way ANOVA test was performed with a Tukey post-hoc test. Significance values are denoted as follows: p< 0.05 *, p< 0.01 **, p< 0.001 ***. Box plots: line, median; box, 75th–25th percentiles; whiskers, minimum to maximum.