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. 2016 Oct 7;7:12972. doi: 10.1038/ncomms12972

Figure 9. Effect of FOXO levels on PIP3 and actomyosin cables.

Figure 9

(a–b) PIP3 accumulation and (c–g) actomyosin cable formation during wound healing in larvae with modified levels of InR signalling and FOXO. (a) tGPH distribution during the early phase of wound healing. (b) Time course of fluorescence changes in wound edges relative to t=0 (set at 100). Mean±s.e.m. one-way ANOVA with post hoc test ***P<0.0001 for control versus A58>InRDN and °P<0.01 control versus A58>foxo, n=4 larvae each genotype. (c) Time-lapse images of actomyosin cable assembly during wound healing in L3 larvae. (d) Average time for initiation of actomyosin cable formation for genotypes shown in c: 6 min for control, 12 min for A58>InRDN, 13 min for A58>foxo-GFP, 6 min for foxoΔ94/Δ94 and 8 min for >InRDN, foxoΔ94. Mean±s.e.m. one-way ANOVA with post hoc test, *P<0.01, **P<0.001 and ***P<0.0001. (e) Kinetics of actomyosin cable contraction. Time course of changes in relative cable circumference. Mean±s.e.m. one-way ANOVA with post hoc test, *P<0.01 (black * marks for control versus A58>InRDN and grey * marks for control versus A58>foxo), °P<0.01, °°P<0.001 and °°°P<0.001 (black ° marks for foxoΔ94/Δ94 versus A58>InRDN and grey ° marks for foxoΔ94/Δ94 or A58>InRDN,foxoΔ94/+ versus A58>foxo. n=4–5 larvae each genotype. (f) Micrographs of actomyosin cables in control and foxoΔ94/Δ94, 28 and 30 min after wounding. (g) Quantification of cable thickness in control and foxoΔ94/Δ94 wounds. For each wound, 6 radial lines were drawn and the fluorescent intensity plotted along a length of 10 μm of the radius, starting at ∼1 μm inside the actin cable. (n=3 wounds for each genotype). Mean±s.e.m. two-tailed Student's t-test, *P<0.05 for distances of 0.9–1.3 μm and 2.9–3.7 μm and ***P<0.0005. for 1.4–2.8 μm. Scale bars, (a,c,f) 20 μm. Transgene genotypes of larvae: (a–b) Control (tGPH/+; A58-Gal4/+), A58>InRDN (tGPH/+ A58-Gal4, UAS-InRDN), A58>foxo (tGPH/+; A58-Gal4, UAS-foxo). (c–g) Controls (Sqh-mCherry/+, A58Gal4/+), A58>InRDN (Sqh-mCherry/+, UAS-InRDN; A58-Gal4), A58>foxo (Sqh-mCherry/+; UAS-foxo-GFP, A58-Gal4), foxoΔ94/Δ94(Sqh-mCherry/+; foxoΔ94/foxoΔ94), >InRDN, foxoΔ94 (Sqh-mCherry/+, UAS-InRDN; A58-Gal4, foxoΔ94/+).