Fig. 3

Limiting dilution cell transplantation shows that myf5 can confer tumor-propagating ability to differentiated myf5:GFP+/mylpfa:mCherry+ cells.

Tumors were generated in myf5:GFP/mylpfa:mCherry CG1-strain syngeneic zebrafish. Representative tumors arising in primary transplanted fish (1°T, A–C) or secondary transplanted fish following engraftment with highly purified myf5:GFP+, mylpfa:mCherry-negative (2°T G+, D–F) or myf5:GFP+, mylpfa:mCherry+ ERMS cells (2°T G+R+, G–I). Sort purity following FACS is noted in the lower left panels of D and G and was >92% for each population. These cells were used for cell transplantations and data provided in D-I. Cell viability was >95%. (J,L) Graphical summary of tumor engraftment following limiting dilution cell transplantation using highly purified sorted ERMS cells. Data is combined from all tumors shown in Table 1. ***p<0.0002 by ELDA analysis. (K,M) Relative gene expression analysis of sorted G+ or G+R+ ERMS cells from representative kRAS^G12D (K) or kRAS^G12D; mylpfa:myf5 (M) expressing ERMS (Standard Deviation, n = 3 technical replicates per PCR condition). *p<0.05; **p<0.01 and ***p<0.001 by Student’s t-test.