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. 2015 Sep 19;6(33):34389–34401. doi: 10.18632/oncotarget.5360

Figure 5. The suppression of CLU levels in CRC cells overexpressing L1 reduces their metastasis to the liver.

Figure 5

A. Ls174T CRC cells stably overexpressing L1, two independently isolated L1-expressing Ls174T clones in which CLU levels were suppressed by shRNA to CLU and control cells expressing the empty vector (as described in Fig. 4B), were injected into the tip of the spleen of nude mice and the formation of macrometastases in the liver was determined after 4 weeks. Arrows point to tumors formed at the site of injection in the spleen and the arrowheads point to large macrometastases in the liver. B. The weight of the excised livers was determined after 4 weeks. C. The levels of L1 and CLU proteins were determined by western blotting of tumor samples from the spleen and liver of mice to verify that the injected cell clones maintained the expression of the stably transfected L1 and the shRNA to CLU. The elevated endogenous CLU in L1 expressing cells was maintained in liver metastases (lanes 4 and 6), but there was no detectable CLU in L1+shRNA to CLU-expressing cells (lanes 8, 10, 12, 14). ***p < 0.001. Error bars: ±S.D.