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

Figure 4. The elevated CLU expression in CRC cells overexpressing L1 is required for conferring increased motility, proliferation and tumorigenesis.

Figure 4

A. The level of endogenous CLU in L1-expressing Ls174T cells was suppressed by shRNA for CLU and independent cell clones displaying decreased CLU, as determined by western blot analysis with anti CLU antibody, were isolated (lanes 3–6). Lane 2 shows a clone with minimal decrease in CLU level after shCLU RNA transfection. B. The colony morphology of clones expressing the empty vector (Con1), L1, and two clones expressing L1+shRNA to CLU (L1+shCLU Cl1 and Cl2). C. The expression of E-cadherin was determined by western blot analysis in the clones described in B. D. The motility of the clones described in A. was determined in a “scratch-wound” experiment as described in Fig. 3C. E. The CRC cell clones described in B. were analyzed in triplicate cultures for their proliferation in 0.1% serum. On day 5, the proliferation of L1-expressing Ls174T cells were statistically different from all other cell clones (p < 0.01). F. The cells described in B. were also injected s.c into each mouse at four different sites in the flanks of nude mice. After 2 weeks the nude mice were sacrificed, the tumors were excised and photographed. G. The weight of the excised tumors was determined. *p < 0.05, **p < 0.01, ***p < 0.001. Error bars: ±S.D.