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. 2020 Oct 2;8:587697. doi: 10.3389/fcell.2020.587697

FIGURE 4.

FIGURE 4

Stromal Per2–/– is essential for the formation of the pre-metastatic niche. WT and Per2–/– mice were injected with MC38 cancer cells into the hepatic portal vein and sacrificed 1 or 3 weeks post injection, as indicated. PBS was injected to control mice which were sacrificed 1 week post injection. n = 4–5 mice per genotype. (A) Representative H&E staining of livers from WT and Per2–/– mice post injection of PBS or MC38 cancer cells. Scale bar – 200 μm, inset – 67 μm; T-tumor, N-normal tissue. (B) The area covered by metastases in MC38-injected livers was calculated by QuPath based on H&E images. P-value was calculated using two way ANOVA. Error bars represent SEM, ***p < 0.0005. (C) RNA-sequencing was performed on livers of WT and Per2–/– mice injected with MC38 (n = 4 per genotype) and PBS (n = 2 per genotype). Heatmap showing hierarchical clustering of 1222 differentially expressed genes between all four conditions. Pathway analysis was performed using Metascape. Significant pathways are shown; p < 0.05, FDR < 0.05 (for details see Supplementary Table 2).