Extended Data Fig. 7: Proteomics measurements suggest the interaction between RUNX1 and TCF-1 is dependent on the L1 domain.
a) Representative immunofluorescence images depicting GFP tagged wild type (WT) TCF-1, ΔL1 mutant TCF-1 and empty vector (EV). DAPI staining of nuclei and overlay images are included (right). Boxplot of granularity of GFP signal in DN3 cells transduced with either EV, WT TCF-1 or ΔL1 fused with GFP. Granularity indicates the percentage of highest intensity elements of 8 pixels subtracted relative to the background (see Methods). Cells with a more granular pattern or punctate localization are indicated by a lower percentage. Center line of box plots represent median granularity, limits represent 1st and 3rd quartiles, whiskers represent maximum and minimum values, data points represent outliers. Cells analyzed per condition EV: n = 189, WT TCF-1: n = 237, ΔL1: n = 190. P values were determined by a two-tailed Mann-Whitney test: *P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.001. Scale bar: 4µm. b) Heatmap indicating the Z score of the log2 normalized abundance of top 100 proteins detected with a higher enrichment between DN3 cells expressing WT TCF-1 and both EV and ΔL1 in mass spectrometry of a TCF-1 immunoprecipitation in DN3 cells. c) Depiction of L1 dependent TCF-1 protein-protein interaction network identified by mass spectrometry of a TCF-1 immunoprecipitation in DN3 cells. Node size and color indicate fold change in log normalized abundance between DN3 cells expressing WT TCF-1 and EV. d) Network terms corresponding to Uniprot keywords are highlighted in the network depicted in c.
