Fig. 4: Mouse models demonstrate the role of lEV HK1 in HCC progression.
a–d, Mice bearing orthotopic xenografts were intravenously injected with lEVs derived from different activated moHSCs indicated. Representative images (scale bar, 1 cm) and weights of orthotopic tumors are indicated (c; n = 8 independent mice). The expression of HK1 and Ki67 is shown (b, d; scale bar, 100 μm; n = 12 fields from three independent tumor tissues). e–h, Representative images (scale bar, 1 cm) and weights of Hepa1-6 orthotopic liver tumors in Hk1f/f and Hk1f/f;Gfap-Cre (e) or Hk1f/f;Lrat-Cre (g) mice are indicated (e, g; n = 8 independent mice). The expression of HK1 and Ki67 in tumors is shown (f, h; scale bar, 100 μm; n = 10 fields from three independent tumor tissues). i,j, Representative images of the livers (scale bar, 1 cm) in Hk1f/f and Hk1f/f;Gfap-Cre mice with DEN/CCl4-induced HCC (n = 8 independent mice) are indicated, and tumor number and large tumor number (Φ > 3 mm) per mouse were quantified (i). The expression of HK1 and Ki67 in tumors is shown (j; scale bar, 100 μm; n = 12 fields from three independent tumor tissues). k–n, Representative images of the liver (scale bar, 1 cm), total tumor number and large tumor number (Φ > 3 mm) per mouse and liver/body weight ratio in Hk1f/f and Hk1f/f;Gfap-Cre mice (k; n = 14 independent mice), or Hk1f/f;Lrat-Cre mice (m; n = 12 independent mice). The expression of HK1 and Ki67 is shown (l, n; scale bar, 100 μm, n = 12 fields from three independent tumor tissues). o,p, lEVs derived from control or HK1 knockdown moHSCs were injected via tail vein into mice that were intravenously inoculated with luciferase-expressing Hepa1-6 cells. Tumor metastases were indicated by the luciferous signals and hematoxylin and eosin (H&E) staining (o; n = 8 independent mice). The HK1 expression in metastatic tumors are shown (p; scale bar, 100 μm. Three independent mice in each group were detected with similar results. Statistical data are presented as the mean ± s.e.m. Statistical analyses were determined by two-tailed Student’s t-test (e–n) and one-way ANOVA, followed by Tukey’s post hoc test (a–d, o).
