Extended Data Fig. 10: Functional interrogation of flies with reduced gut tracheation.
a, Expression of bnlRNAi from gut muscles (Hand>bnlRNAi) does not change mitotic indices in midgut of 7-day-old flies seen by number of pH3-positive cells. b, Hand>bnlRNAi expression marginally reduces total number of sips during 1 h of feeding, as recorded in a FlyPad device (see Methods). c-d, Hand>bnlRNAi expression does not alter FCF blue-dyed food contents in gut of adult flies fed ad libitum (c) or starved for 16 h and re-fed for 15 min (d), as measured by blue dye absorbance at 634 nm from cleared homogenates of groups of 3 flies. e, Hand>bnlRNAi expression does not alter food transit time in gut, measured by percentage of flies with dyed food present in portions of the gut (midgut, hindgut and ampulla). Only size-matched guts are shown to rule out effects of gut size (see Methods). f, Hand>bnlRNAi expression does not alter the total number of faecal deposits, quantified by bromophenol blue-dyed deposits after 60 h of feeding (see Methods). g-h, Hand>bnlRNAi expression reduces survival in male (g) and female (h) flies starved in 1% agar relative to respective controls, shown by Kaplan-Meier survival curves. Boxplots: line = median, box = first quartile and third quartile, whiskers = minimum and maximum. n = number of biologically independent samples. Statistical significance was assessed using one-way ANOVA followed by Tukey post-hoc tests (a–d,f), logistic regression with Chi2 post-hoc tests (e) and log-rank test (g,h): non-significant (n.s.) = p > 0.05; * = 0.05>p > 0.01; ** = 0.01>p > 0.001; *** = 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).
