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. 2022 Feb 2;13:626. doi: 10.1038/s41467-022-28076-3

Fig. 5. Spontaneously metastasizing tumor cells are more frequently doubly positive for dormancy and stem-like markers compared to intravenously injected tumor cells.

Fig. 5

a Representative images of triple immunofluorescence staining for GFP, NR2F1, and SOX9 expression in primary tumors, circulating tumor cells (CTCs), and disseminated tumor cells (Lung) from an E0771-GFP SM model (Left) and in disseminated tumor cells (Lung) from an EM model (Right). Green = GFP; Red = NR2F1; Orange = SOX9; Blue = DAPI. Scale bar for Primary Tumor = 50 μm. Scale bar for CTCs and Lung = 15 μm. b Percentage of double-positive tumor cells NR2F1-positive SOX9High from each group in Fig. 5a. Primary Tumor: n = 2383 in 97 fields of view (65 × 65 µm2) in 7 animals; CTCs: n = 379 cells in 8 animals; SM Lung: n = 104 cells in 9 animals; In vitro: n = 413 cells in 3 independent experiments. EM Lung: n = 75 cells in 7 animals. Bar = mean. Error bars = ±SEM. For EM Lung vs. SM Lung (p = 0.0001) and EM Lung vs. in vitro (p = 0.69), a two-tailed Kruskal-Wallis test with Dunn’s multiple comparisons adjustment was used. For PT vs. CTC: (p = 0.0041), PT vs. Lung SM (p = 0.0030), and CTC vs. Lung SM (p = 1.00) a two-tailed ANOVA test with Sidak’s multiple comparisons adjustment was used. **p < 0.01. ***p < 0.001. ns = not significant. Source data are provided as a Source Data file.