Figure 4.

SPNS2 affected ID-enhanced HCC metastasis in an immune-independent manner in vivo

(A and B) Nude mice were fed with either IA or ID diet at day −7. Human HCC Huh7 cells were orthotopically or intravenously administrated at day 0. All mice were sacrificed at day 50. Representative figures were taken at day 50. n = 3. (A) Growth of tumor cells over time in the lung of mice. (B) N and T tissues in the liver were sectioned from the orthotopically administrated mice. mRNA and protein expression of SPNS2 and iron-related genes were determined. (C and D) Growth of tumor cells over time in the lung of mice. (C) NCG mice were fed with either IA or ID diet at day −7. Human HCC Huh7 cells (upper) or mouse HCC H22 cells (lower) were orthotopically administrated at day 0. (D) NCG mice were fed with IA diet at day −7. TFRC-knockdown Huh7 cells (upper) or H22 cells (lower) and their control counterparts were orthotopically administrated at day 0. Representative figures were taken at day 50. Data were presented as mean ± SEM. ∗p < 0.05, ∗∗p < 0.01 versus IA diet, or versus shNC.