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. 2016 Feb 17;6:21596. doi: 10.1038/srep21596

Figure 7. Knockdown of USP4 inhibited brain metastasis and promoted overall survival (OS) and brain metastasis-free survival (BMFS).

Figure 7

(A) PC14PE6/LvBr5-Luc cells were transfected with control (CTRL) siRNA or USP4-specific siRNA for 48 h. The levels of USP4, β-catenin, E-cadherin, and ZEB1 were determined by western blotting. (B) For the invasion assay, equal numbers of transfected cells were inoculated into a Transwell and invaded cells were stained and counted under a microscope. Data are means and standard deviation from more than three independent experiments. (C) To investigate the effect of USP4 silencing on brain metastasis in vivo, transfected PC14PE6/LvBr5-Luc cells were directly injected into the left ventricle (LV) of the heart. Bioluminescence images were acquired at 22 days post-injection. (D,E) Incidence of whole body (D) and brain metastasis (E) were quantified by the luminescent signal at a given time point. (F,G) Kaplan–Meier curves and p-values for overall survival (OS) and brain metastasis-free survival (BMFS) were analyzed in systemic metastasis models. *p < 0.05.