Fig. 3: Patterning competency is specific to undifferentiated blastema tissue.

a Experimental set-up to analyze the expressional responses of different-staged amputation blastemas when treated with RA or DMSO (early bud (EB), apical late bud (Ap. LB), basal LB (Ba. LB), and mature (Mat.) tissue treated with either RA or DMSO via qRT-PCR. Red lines indicate the locations of amputations. Blue line represents the border between the basal and apical regions of the LB blastema. b Histograms representing the qRT-PCR data of Shh, Alx4, Fgf8, and Meis2 (normalized to Ef1a expression) in treated mature (Mat.), EB, apical-LB (Ap. LB), and basal-LB (Ba.-LB) blastemas (N = 4–6 biological replicates, using the Unpaired t-test relative to EB, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, error bars are SEM). c Description of CALM-P treatment scheme and sample generation following an upper-arm denervation surgery (red line) or sham surgery. d Histogram representing the qRT-PCR data of the relative expression of Shh (normalized to Ef1a expression) in the CALM-P samples described in “C”. “S” indicates sham surgery samples, “D” indicates denervated samples. (N = 5 biological replicates, using the Unpaired t-test, *p < 0.05, error bars are SEM). e Description of near-wound denervated (red line) CALM-A treatment scheme and sample generation. f Histogram representing the % total of CALM-A surgeries that led to different ectopic growth responses in sham (S) and near-wound denervated (D) CALM-A blastemas treated with RA (+) or DMSO (−) (N = 20 biological replicates per sample—refer to Supplementary Data 1.1).