FIG 5.

USP6 promotes cell migration and invasion. (A) HeLa cells were infected with the indicated lentivirus to generate stable cell lines, and 2 × 10⁴ HeLa cells were seeded into the upper chamber of each Transwell. Twenty-four hours later, the cells were subjected to transwell assays to measure cell migration. (B) Approximately 2 × 10⁴ of the indicated HeLa cells were seeded into the upper chamber of a Transwell containing Matrigel. Twenty-four hours later, the cells were subjected to an in vitro invasion assay. A zymography assay was performed to determine MMP activity as described in Materials and Methods. (C) HeLa cells were infected with lentivirus carrying shGFP or USP6 shRNAs to generate stable cell lines, and 2 × 10⁴ of the HeLa cells were seeded into the upper chamber of a Transwell containing Matrigel. Thirty-six hours later, the cells were subjected to an in vitro invasion assay. (D) The indicated HeLa stable cell lines were infected with lentivirus carrying shGFP or c-Jun shRNAs; 48 h later, approximately 2 × 10⁴ of the HeLa cells were seeded into the upper chamber of a Transwell containing Matrigel. Thirty-six hours later, the cells were subjected to an in vitro invasion assay. All experiments were performed 3 times. In all panels data are presented as means and SEM for three triplicate samples and were analyzed by t test. *, P < 0.05.