Fig. 6: Tig1 reprogrammes distal cells to proximal identities.

a Experimental design. Tig1-T2a-GFP or Tig1^P155A-T2a-GFP were electroporated in the distal, mCherry⁺ domain of late bud blastemas of Hoxa13:mCherry^Etnka axolotls; FACS isolation followed by bulk RNAseq analysis was carried out for the indicated cells at 3 or 8dpe. b tSNE plot using the first and second dimension shows consistent behaviour between replicates and separation between treatments. c Heatmap of the 2000 most differentially expressed genes between conditions. d GO enrichment analysis of Biological Process for three gene groups determined by K-means hierarchical clustering, displaying the top ten enriched terms for cluster A. The enriched pathways are listed with their corresponding adjusted p values and the number of genes. The p values are corrected for multiple testing using a false discovery rate (FDR). e–h Tig1 overexpression in distal cells promotes a transcriptome shift toward proximal identity. Bars represent ratios of gene expression in Tig1 overexpressing cells versus control cells. e Relative gene expression changes in PD markers. Note that Tig1 overexpression leads to the upregulation of proximal factors Meis1a and Prod1 and the downregulation of distal genes including the master regulator Hoxa13 (e) and its downstream network (f). mCherry protein persists in distal cells even if Hoxa13 is downregulated. g Relative gene expression changes in genes associated with blastema growth. h Relative gene expression changes in cell adhesion/matrix remodelling factors. Adj. p values: *p < 0.05; **p < 0.001. Source data (including exact Adj. p values for e–h) are provided as a Source Data file.