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. 2010 Jan 19;102(4):774–782. doi: 10.1038/sj.bjc.6605536

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Copyright © 2010 Cancer Research UK

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Expression of a full-length CLCN4 induces colon cancer cell migration. (A) A pool of RKO colon cancer cells stably expressing the indicated constructs were lysed and 20 μg protein was analysed by western blotting using a rabbit anti-HA tag antibody (Invitrogen cat no. 715500), followed by an HRP-coupled goat anti-rabbit IgG. Immunoreactive proteins were visualised by enhanced chemiluminescence. Size markers are indicated. (B) A pool of RKO clones overexpressing wild-type CLCN4 (RKO-CLCN4) or the empty vector (RKO (Vector)) were assayed for migration as described in Figure 2 and using 10% FBS as chemoattractant. (C) In vitro invasion was as described for the migration assays, with the exception that a Matrigel-coated filter was used and cells traversing the coated filter were enumerated 16 h later. The experiment was repeated three times and data are shown as average±s.e values. For (B) and (C), an asterisk indicates P<0.05, in comparison with the RKO-CLCN4 data. (D) RT–PCR was carried out for CLCN4 expression as described in Figure 3.