Extended Data Fig. 8: Genetic sex reversals of intestinal muscles.
a-c, Hand>traRNAi.JF03123 masculinizes bnl expression at 5 h after pupal eclosion (APE), seen by bnl-lexA>myr::GFP (top (a), quantified in (b)) and reduces tracheal branches (QF6>mtdTomato) (middle (a), quantified in left (c)) and the number of DSRF-positive nuclei (bottom (a), quantified in right (c)). d, PCA plot of Hand>traRNAi gut shape variability. Females are significantly different in shape to controls (Supplementary Tables 32-33). e, Average Hand>traRNAi gut centrelines show altered shape in the central midgut region in females relative to controls. f, Multidimensional scaling plot representing similarity in gut curvature normalized by gut length between Hand>traRNAi and controls. Arrows represent shift in the mean from control to Hand>traRNAi. g, Hand>traRNAi females have reduced average gut curvature normalized by gut length along midgut loop region relative to controls. h, PCA plot of Hand>SxlRNAi gut shape variability. Groups are significantly different in shape (Supplementary Tables 34-35). i, Hand>SxlRNAi guts show differences in length in females relative to controls. j, PCA plot of Hand>Sxl gut shape variability. Males are significantly different in shape to controls (Supplementary Tables 36-37). k, Hand>Sxl guts show no differences in length relative to controls. Line graphs: mean and standard deviation. Boxplots: line = median, box = first quartile and third quartile, whiskers = minimum and maximum. d,f,h,j, Ellipses represent 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 (c,i,k): non-significant (n.s.) = p > 0.05; * = 0.05>p > 0.01; *** = 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). Scale bars: 200 μm.
