Fig. 5

a Panther Pathways significantly overrepresented by genes repressed by 2-DG in the blastema (Cluster 2). b Violin plots of tgfb1, inhbaa, inhbb, tgfbr2b, snai1a, and hmga2 expression in control and 2-DG treated blastemas (cluster 2). c Image of the tail of a 96 HPA embryo treated with DMSO or the Alk4, 5, and 7 inhibitor SB431542. Drug was added to embryo media 2 h prior to amputations and treatment persisted through the duration of experiment. Scale bar = 200 µm. d Quantification of tail area of 96 HPA embryos treated with DMSO or SB431542. Drug was added to embryo media 2 h prior to amputations and treatment persisted throughout the duration of experiment. Mean and SEM are shown. N = 13 for each condition. Statistics were determined with an unpaired two-tailed t-test. e In situ hybridization of snai1a and msx3 in DMSO, 2-DG, or SB431542 treated 48 HPA embryos. Scale bar = 100 µm. f Stacked percentage graph of snai1a and msx3 expression as determined by in situ hybridization. A score of 1 represents little to no staining, while a score of 4 represents strong staining. N = 10 embryos for all conditions. g Immunofluorescent staining of P-SMAD2 in uninjured, 24 HPA, and 48 HPA control or 2-D treated embryos. Images are sum z-stack projections of 1 µM slices through the entire embryo. B indicates predicted blastema. Scale bar = 50 µm. h Quantification of P-Smad2 intensity in the blastema. Mean and SEM are shown. N = 10 embryos for all conditions. Statistics were determined with an unpaired two-tailed t-test.