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. Author manuscript; available in PMC: 2019 Jan 8.
Published in final edited form as: Cancer Cell. 2018 Jan 8;33(1):108–124.e5. doi: 10.1016/j.ccell.2017.12.001

Figure 6. Purified tumor cells retain expression of myogenic specification factors critical for head and neck muscle development.

Figure 6

(A) FACS isolation of PECAM1 and Tomato stained cells from AST SCM and tumors.

(B) Sphere formation of tumor cell populations sorted as in (A)(n = 4). Scale bars are 100 μm.

(C–E) Expression of SmoM2-YFP and Shh target genes (C), myogenic markers (D) and endothelial markers (E) in populations isolated as in (A), mature SCM from SmoM2/M2 mice and whole tumor from AST mice. Data shown are normalized to Actb expression and expressed relative to sorted Tom−PECAM1− cells (n = 3, mean ± SEM).

(F) Gene ontology analysis of genes upregulated in Tom+PECAM1− cells versus Tom+PECAM1− cells isolated from AST tumors.

(G) Real-time PCR of myogenic specification factors in cells isolated in (A). Data are analyzed as in (C)(n = 3, mean ± SEM).

(H) Representative dual ISH for Tbx1 and IHC for Tomato in sagittal section of E15.5 AST embryo. Right panel is enlarged boxed region on left with high magnification inset shown (n = 4). Scale bars, 100 μm (left) and 20 μm (right).

(I) Representative immunostaining for Tomato (red), MYOD1 (green), and DAPI (blue) in sagittal sections of E15.5 AST embryo shown in H. (n = 4). Scale bar, 50 μm.

*p < 0.05, ** p < 0.01, *** p < 0.001.

See also Figure S6 and Tables S1 and S2.