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Fig. 5 | Genome Biology

Fig. 5

From: A comprehensive enhancer screen identifies TRAM2 as a key and novel mediator of YAP oncogenesis

Fig. 5

TRAM2 stimulates cell migration and invasion. a, b In vitro migration assays of MCF10A and MDA-MB-231 cells transduced with the indicated lentiviral vectors. Cells were labeled with GFP for tracking cells under the microscope. Error bars indicate SD from counted cells in each condition (for a: Control: n = 16962; TRAM2: n = 5095; YAP: n = 17421; for b: Control: n = 18448; TRAM2: n = 8219). ***P < 0.005, two-tailed Student’s t test. c In vivo migration assays of GFP-labeled MCF10A cells transduced with the indicated lentiviral vectors. Cells were injected into the fat pad of NOD-scid IL2Rgammanull female mice, and 14 days after transplantation cells were tracked by intravital imaging for at least 6 h. In vivo migration speed was tracked for cells that could be followed for a minimum of 4 h. Error bars indicate SEM from 1119 control cells (N = 4 tumors), 680 TRAM2 cells (N = 4 tumors), and 1746 YAP cells (N = 4 tumors) respectively. ***P < 0.005, Welch-corrected two-tailed Student’s t test. d Representative overview images of Matrigel-coupled wound-healing invasion assays of MCF10A cells transduced with the indicated vectors at 0 and 72 h. e Wound confluence was calculated using IncuCyte system based on 4 independent Matrigel-coupled wound-healing invasion assay experiments, as in d. f Representative overview images of mammary tumors after injection of MCF10A, MCF10A-YAP5SA, and MCF10-TRAM2 cells (GFP labeled) in the mammary fat pad of female mice 14 days and 35 days after transplantation. Shown are 3D (left-hand panels) and single cross section images (right-hand panels). Purple, collagen; green, GFP fluorescence from injected labeled cells. g MCF10A cells were transduced with the indicated lentiviral vectors, and injected into the mammary fat pad of female NOD-scid IL2Rgammanull mice. Tumor growth kinetics were measured using an external caliper. Error bars indicate SD (N = 4 tumors per condition)

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