CLIC1 supports invadopodia formation in vitro and metastasis in vivo.

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CLIC1 supports invadopodia formation in vitro and metastasis in vivo. CLIC1 supports invadopodia formation in vitro and metastasis in vivo. A–D, 786-0, HT1080, and endothelial cells (HUVEC) were analyzed for invadopodia formation 24 hours after embedding in a 3D fibrin matrix by phase contrast microscopy. A, representative images of fibrin-embedded HUVEC, 786-0, and HT1080 cells are shown (top, controls; bottom, CLIC1-knockdown cells). Scale bars, 50 μm. B–C, phase contrast microscopy images of fibrin clots were analyzed for percentage invadopodia-positive 786-0 and HT1080 cells (B) and overall cell count (C) per optical field after CLIC1 silencing with two CLIC1 shRNA vectors (shCLIC1-2 and 4) compared with scrambled shRNA (shSCR). D, invadopodia formation in fibrin-embedded HUVEC after transfection with CLIC1 siRNA (control, nontargeted siRNA) or after treatment with 200 μmol/L IAA94 (control, DMSO). E, the number of lung tumors (tumor multiplicity) was assessed 3 weeks after i.v. injection of 5 × 105 CLIC1 or SCR shRNA-transformed HT1080 cells. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Lisa A. Gurski et al. Mol Cancer Res 2015;13:273-280 ©2015 by American Association for Cancer Research