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

Fig. 4. Spontaneously metastasizing tumor cells are more frequently positive for dormancy or stem-like markers compared to intravenously injected tumor cells.

Fig. 4

a Representative immunofluorescence images of NR2F1 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, Blue = DAPI. Scale bar for Primary Tumor = 50 μm. Scale bar for CTCs and Lung = 15 μm. b Percentage of NR2F1-positive and negative tumor cells in each group in A. Primary Tumor: n = 2,041 cells in 115 fields of view (65 × 65 µm2) in 8 animals; CTCs: n = 550 cells in 8 animals; SM Lung: n = 237 cells in 12 animals; EM Lung: n = 199 cells in 8 animals. In vitro: n = 463 cells in 5 independent experiments. Bar = mean. Error bars = ±SEM. For all comparisons, a two-tailed one-way ANOVA test with Sidak’s multiple comparisons adjustment was used (PT vs. CTC: p < 0.0001; PT vs. Lung SM: p < 0.0001; CTC vs. Lung SM: p = 0.80; Lung SM vs. Lung EM: p < 0.0001; In vitro vs. Lung EM: p = 1.00). ****p < 0.0001. ns = not significant. c Representative immunofluorescence images of 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 = SOX9, Blue = DAPI. Scale bar for Primary Tumor = 50 μm. Scale bar for CTCs and Lung = 15 μm. d Percentage of SOX9High tumor cells from each group in C. Primary Tumor: n = 4,633 cells in 150 fields of view (65 × 65 µm2) in 8 animals; CTCs: n = 558 cells in 5 animals, SM Lung: n = 341 cells in 11 animals; EM Lung: n = 182 cells in 8 animals. In vitro: n = 298 cells in 3 independent experiments. Bar = mean. Error bars = ±SEM. For CTC vs. Lung SM (p = 0.89) a two-tailed one-way ANOVA test with Sidak’s multiple comparisons adjustment was used. For PT vs. CTC: (p = 0.0027), PT vs. Lung SM (p = 0.0002), In vitro vs. Lung EM (p = 1.00), and Lung SM vs. Lung EM (p = 0.0011) a two-tailed Kruskal-Wallis test with Dunn’s multiple comparisons adjustment was used. **p < 0.01. ***p < 0.001. ns = not significant. Source data are provided as a Source Data file.