Fig. 4: Dilp8 is critical for progression of the pupariation motor program.

a Muscle calcium (mhc»GCaMP) fluctuations of a single WT (dilp8 +/−) larva (whole-body measurement, blue). Pupariation motor program (PMP). b Speed (black), and distance traveled by (red) the same larva depicted in a. c PMP in (a) and its specific stages. Shown are mhc»GCaMP (blue) and aspect ratio (AR-GCaMP, green) fluctuations. d Scheme describing the parameters measured for the pre-GSB contractions. e Dot plots showing the number of pre-GSB contractions of WT (dilp8+/−) and dilp8 mutant (dilp8−/−) animals. f–h Dot plots showing the average f duration, g amplitude, and h period of pre-GSB contractions in WT and dilp8 mutants. i Time-lapse of GCaMP oscillations during a WT pre-GSB contraction. Anteriormost segments are initially extruded (arrowhead) by the strong whole-body contraction and subsequently internalized by the activation of ventral longitudinal muscles (arrows). Representative profile from 3 recorded animals. j An example of muscle calcium (mhc»GCaMP) fluctuation (blue) and aspect ratio (AR-GCaMP, green) fluctuations of a dilp8 mutant animal that showed pre-GSB-like contractions and one that k did not show any detectable pre-GSB contractions. l dilp8 mutants fail to increase the duration of the pre-GSB contractions with time. Shown are dot plots of the duration of the first and last two pre-GSB contractions of WT and dilp8 mutants. Statistics (full details in Supplementary Table 2): e–h, l Dot: average per larva. Horizontal bar, median. Error bars: 25-75%. e, g, h Mann–Whitney Rank sum test. f Student’s t-test. l Dunn’s test. Same blue letters, P > 0.05. *P < 0.05. P = 0.76 in e (excluding animals with no contractions). (N) Number of animals (orange). Scale bar, 1 mm.