Fig. 7

GPCR inhibition disrupts islet assembly. (A) Transgene hsp70:LifeActTom-PTX was used to inducibly express pertussis toxin (PTX, top). The transgene hsp70:LifeActTom served as a control (bottom). (B) Scheme of islet assembly experiment with heat shock induction of PTX. (C) Islet volume quantitation of heat shock-treated controls (n=12, 99 objects), compared with larvae with induced expression of LifeActTom (n=14, 120 objects), or LifeActTom-PTX (n=6, 61 objects). Images were captured on a Zeiss LSM5 and volume quantitation performed using ImageJ (minimum object size 100 µm³). *P<0.05, **P<0.01, Kruskal–Wallis test followed by Dunn's multiple comparison test. ns, not significant. Results are representative of two independent experiments. (D,E) Selected confocal projections from time-lapse series of control (D) and PTX-expressing (E) transgenic larvae at 7 dpf, 4 or more hours following a heat shock. Scale bars: 10 μm. (F,G) Quantitation of cell solidity (F) and area (G) measured in individual frames of time-lapse series with images acquired at 4 min intervals (as shown in Fig. S16A,B). ***P<0.0001, Mann–Whitney test, two-tailed. Data are combined from a total of eight control and eight PTX-expressing larvae analyzed in three separate experiments. Control samples include non-heat-shocked hsp70:PTX transgenics, and LifeActTom⁺ heat-shocked samples. Additional sample information is provided in Table S5.