Extended Data Fig. 1: Sex differences, variation and plasticity of gut shape.
a,b, Sample layout for microCT, showing parallel stacking of sample tubes (a) and a radiograph exemplifying a batch scan of multiple Drosophila specimens (b). c, PCA plot of shape variability in hindgut centrelines for wild-type OregonR. Male and female hindguts are significantly different in shape (Supplementary Table 5-6). d-f, Relative length (% of whole gut length) (d) and absolute length (e) of different gut regions and average gut radius (f) for OregonR. g-h, Gut torsion normalized by length (g) and radius (h) along midgut loop region for OregonR. i, PCA plot of wild-type CantonS whole gut shape variability showing male and female guts are significantly different in shape. j,k, Average CantonS gut centrelines (j) and gut length (k) showing differences between males and females. l, PCA plot of w1118 gut shape variability showing male and female guts are significantly different in shape. m,n, Average w1118 gut centrelines (m) and gut length (n) showing differences between males and females. o, Anteroposterior slices of female microCT scans overlaid with 3D reconstructions of the gut (orange) and ovaries (brown), showing differences in gut position between ovoD1 mutant females and controls. p, PCA plot of ovoD1 gut shape variability showing significant difference in shape to controls. q,r, Average ovoD1 mutant female gut centreline (q) and gut length (r) showing differences relative to controls. s, Anteroposterior slices of female microCT scans overlaid with 3D reconstructions of the gut (orange) and ovaries (brown), showing gut and ovary positions in flies starved for 48 h and controls. t, PCA plot of 48 hour-starved gut shape variability showing significant difference in shape to controls. u,v, Average gut centrelines (u) and gut length (v) of flies starved for 48 h, showing differences relative to controls. Line graphs: mean and standard deviation. Boxplots: line = median, box = first quartile and third quartile, whiskers = minimum and maximum. PCA plots: ellipses represent the 95% confidence space for each group. Diagrams represent the extremes of variation along each PC (see methods). n = number of biologically independent samples. Statistical significance was assessed using one-way ANOVA followed by Tukey post-hoc tests (d,e,r,v) or a two-sided two-sample t-test (f,k,n): non-significant (n.s.)= P > 0.05; ***= P < 0.001. See Supplementary Information for exact P-values, statistical tests and sample sizes. Males are in blue, females in orange and controls in lighter matching colours. Ctrl = control group (see genotypes in Supplementary Information).
