Fig. 3

Hedgehog signalling plays a central role during larval tail regeneration. a Continuous treatment with the Hedgehog pathway inhibitors cyclopamine (20 μM) and LDE225(20 μM) blocks regeneration (Cyclopamine 10/10, LDE 10/10, number of experiments = 2) (Control 20/20, number of experiments = 4). Cyclopamine pulse treatment blocks regeneration (8/10, number of experiments = 2). b Larvae treated with cyclopamine (20 μM) and LDE225 (20 μM) from 0 to 48 hpe have reduced expression of dlx5a and msxc (for all panels results are 10/10, number of experiments = 2). c Wound-induced expression of markers for the Wnt/β-Catenin, FGF and RA pathways is lost after treatment with cyclopamine. Larvae in the wnt10a and pea3 panels were treated with 50 μM cyclopamine from 0 to 30 hpe. tcf7, fgf10a and raldh2 panels were treated with 20 μM cyclopamine from 0 to 48 hpe (for all panels results are 10/10, number of experiments = 2). d Cell proliferation is inhibited by treatment with cyclopamine (control n = 10, cyclopamine n = 8, number of experiments = 2). This analysis was done as in Fig. 1d. Unpaired t test two-tailed results in P < 0.0001 indicated as ****. e Upregulation of Wnt/β-Catenin signalling after cyclopamine treatment restores expression of dlx5a, msxc and raldh2 (for all panels results are 10/10, number of experiments = 3). Arrowheads indicate partial rescue of marker expression. Larvae were treated from 0 to 40 hpe with 50 μM cyclopamine and then incubated in 10 μM GskXV until fixation