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. 2015 Jan 23;17(1):124–133. doi: 10.1016/j.neo.2014.11.009

Figure 6.

Figure 6

Mig6 depletion induces apoptosis and prevents pulmonary metastasis of MDA-MB-231 cells. (A) MDA-MB-231 cells stably expressing a scrambled (sc) shRNA or three independent Mig6 targeted shRNAs were analyzed for Mig6 and EGFR expression by immunoblot. Actin served as a loading control. (B and C) Luciferase expressing, Mig6-depleted MDA-MB-231 cells were grown under 3D culture conditions and cellular outgrowth was quantified by bioluminescence. Photomicrographs in panel B are representative structures formed by control (scram) and Mig6-depleted (shMig6) cells. Arrows indicate apoptotic cell morphologies. Data in panel C are the average bioluminescent (± SE) values normalized to values measured immediately after plating (T0) from two independent experiments carried out in triplicate. (D) The floating cell fraction of control and Mig6-depleted MDA-MB-231 cells were collected and assayed for Caspase 3/7 activity. (E) Control (scram) and Mig6-depleted (shMig6) MDA-MB-231 cells were injected into the lateral tail vein of nu/nu mice. Longitudinal bioluminescent images from the same mice are shown immediately following injection (T0) and at the indicated time points thereafter. (F) Data are the mean (± SE, n = 5 mice per group) pulmonary bioluminescent values, normalized to the injected value (T0). (G) Representative ex vivo lungs isolated from mice 49 days after injection with control (scram) or Mig6-depleted (shMig6) MDA-MB-231 cells.