Extended Data Fig. 7: Tumor-propagating potential is enriched in the mesenchymal-enriched tumor cell fraction in FN-RMS.
a,e,i,m, Flow cytometry analysis of FN-MSK74711 cells harvested directly from PDX tumors grown in NSG mouse (a) or cell line models (e,i,m) prior to (left) and after FACS enrichment (right two panels). b,f,j,n, Quantitative PCR confirming cell state enrichment following FACS (n = 2 independently engrafted tumors (MSK74711) and analyzed in triplicate from independent sorts, 6 datum points shown). Three replicates were analyzed from independent sorting of RD, 381 T and SMS-CTR cells. Two-way ANOVA followed by two-sided Student’s t-test comparison (*P < 0.05, **P < 0.01; ***P < 0.001 and ****P < 0.0001). c,g,k,o, Representative images of sphere size following FACS enrichment and plating for two weeks (scale bar = 20 μm). Shown is a representative micrograph of tumorspheres (left) with similar results seen across replicate wells. Combined quantification from two independent experiments (c,g,k) and three independent experiments in panel o. Three wells/experiment were completed and the combined analysis shown (o, +/− SEM noted). d,h,l,p, Quantification of tumor sphere numbers from representative limiting dilution experiments (datum points represent number of spheres found within individual wells from a single experiment). Experiments replicated twice (d,h,l) or three times (p) with similar results. Mean + /− SEM noted for SMS-CTR, two-way ANOVA followed by two-sided Student’s t-test (*P < 0.05, **P < 0.01, ***P < 0.001). Mesenchymal-enriched (Mesen, Mes or Me, muscle (Musc, Mu), interferon (INF), proliferative (Prolif).
