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. 2023 Feb 8;9(6):eade9238. doi: 10.1126/sciadv.ade9238

Fig. 4. PAX3::FOXO1 down-regulation leads to MuSC-like and differentiated subpopulations.

Fig. 4.

(A) Schematic workflow. (B) Representative WB of Rh4 and KFR cells cultured with (+DOX) or without (−DOX) DOX for 48 hours. (C) UMAP plot of 1978 Rh4 and 2589 KFR cells following KD of PAX3::FOXO1. Lines with shP3F1 were cultured with DOX for 48 hours (+DOX) to induce protein down-regulation and profiled by scRNAseq. Control lines that were not exposed to DOX are also shown (−DOX). UMAP plots are colored by DOX exposure (left) or the overall expression (color scale) of the identified signatures (right). (D) Proposed model of PAX3::FOXO1 heterogeneity across aRMS cell lines. Upon PAX3::FOXO1 removal, the differentiation block is released and the oncogenic loop is disrupted. The cycling progenitor subpopulation disappears, and the remaining cells display MuSC-like or differentiated features.