Fig. 4
Timeless accelerates the growth and metastasis of CRC cells in vivo. (a) Expression of Timeless was detected in Timeless-knockdown stable HCT116 cells by western blotting. (b) Images of xenograft tumors from groups of BALB/c-nude mice 4 weeks after the subcutaneous injection of stable Timeless-knockdown HCT116 cells (sh-Tim) or control HCT116 cells (sh-Ctrl). (c) Tumor size was calculated using the equation volume = (length × width2)/2 every 2–3 days 1 week after injection. (d) Weights of xenograft tumors were measured. (e) Hematoxylin and eosin staining and immunohistochemistry for Timeless and PCNA in xenograft tumors. Original magnification, ×200. (f) Western blotting analysis of EMT-related Snail, E-cadherin, and N-cadherin in xenograft tumors. (g) Photos of tumor metastasis in lung tissues (left panel). The number of metastatic foci in lung tissues is quantified in the right. The white arrows show the metastases. (h) Hematoxylin and eosin staining of lung tissues. Original magnification, ×100. Error bars indicate the means ± SD. *P < 0.05, **P < 0.01