Figure 1: miR-135b is upregulated in a highly invasive lung cancer cell line and modulates cell invasion and migration ability.

(a) Real-time RT–PCR was conducted to quantify the endogenous expression of miR-135b, miR-21 and miR-126* in CL-series cell lines. Assays were performed in triplicate, and the results are presented as the fold-change in expression compared with CL1-0. The expression of U6B was used as a normalization control. (b) Migration assays were performed in CL1-0 cells transduced with control (Neo) or miR-135b-expressing (miR-135b) lentiviral vectors. The number of migrating cells in three different fields was counted 15 and 24 h after the inserts were removed. *P<0.05; **P<0.005 by Student’s t-test. (c) An invasion assay was performed in lentiviral vector-modified CL1-0 cells as described in (b). The number of invading cells was counted 20 h after cell seeding and is presented as the mean±s.d., (d, e) CL1-5-F4 and Hop-62 cells were transfected with scramble (NC) or miR-135b-specific antagomir (Antago-135b) for 48 h and then subjected to (Fig. 1d) migration assays and (Fig. 1e) invasion assays as described above. *P<0.05; **P<0.005 by Student’s t-test. (f) CL1-0 cells transduced with control (Neo) or miR-135b-expressing (miR-135b) lentiviral vectors were subjected to a soft agar assay. **P<0.005 by Student’s t-test. (g) CL1-5-F4 and Hop-62 cells were transfected with the negative control (NC) or the Antago-135b for 48 h and subjected to soft agar assays. Error bars indicate mean±s.d., *P<0.05 by Student’s t-test.