(A) In vitro BBB transmigration activity of the 231-Br cells harboring control or Lnc-BM shRNAs. The number of transmigrated cells relative to the control cells is plotted (n = 3 independent experiments, unpaired Student’s t test). Scale bars: 100 μm. (B) Bioluminescence imaging (BLI) (n = 5 animals) of mouse, 4 weeks after intracardiac injection of 231-Br cells harboring indicated shRNAs (1-way ANOVA). Scale bars: 200 μm. White arrows: brain blood vessels. (C) Representative images and statistical analysis of H&E staining of mouse brain tumor burden at 35 days after intracardiac injection of 231-Br cells harboring indicated shRNAs (1-way ANOVA). Scale bars: 200 μm. n = 5 animals per group, 3 sections per brain. Micromet., micrometastatic lesions; Macromet., macrometastatic lesions. (D) Representative images of BLI (n = 8 animals), brain ex vivo bright field and ex vivo GFP, and statistical analysis of brain area photo flux (right panel) 5 weeks after intraarterial injection of HCC1954-Br cells stably expressing indicated shRNAs (1-way ANOVA). White arrow: brain metastatic lesions. Scale bars: 3 mm. (E) Kaplan-Meier plot of survival in the experiment of D (n = 8 animals per group, log rank test). Data are mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001.