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. Author manuscript; available in PMC: 2015 Nov 20.
Published in final edited form as: Cell Rep. 2014 Nov 6;9(4):1318–1332. doi: 10.1016/j.celrep.2014.10.011

Figure 6. The STP-REST axis is required for primary tumor growth and metastatic expansion.

Figure 6

  1. MDA-MB231-LM2 TNBC cells were engineered to inducibly co-express SCYL1 and TEX14 shRNAs. Protein levels were analyzed by western blot.
  2. Depletion of SCYL1 and TEX14 impairs anchorage-independent growth. Cells from A were assessed for anchorage-independent growth. Data presented as mean +/− SE.
  3. In vivo barcode-based competition assay schematic. MDA-MB231-LM2 cells expressing inducible negative control shRNA or combination-shTEX14+shSCYL1 were mixed at an equal ratio. This population was transplanted into mice, and tumors were allowed to form +/−dox. At the experimental end point, genomic DNA was collected, and the relative abundance of each cell population was quantified via barcode-qPCR-analysis.
  4. Depletion of SCYL1 and TEX14 impairs primary tumor growth. The experiment was carried out as described in C. Barcode levels +/−dox are shown. Each dot represents an individual tumor. The black line represents the median.
  5. Depletion of SCYL1 and TEX14 impairs metastatic progression. MDA-MB231-LM2 TNBC cells engineered to inducibly express SCYL1+TEX14 shRNAs or negative control shRNAs were mixed in equal ratio, intravenously injected, and monitored for lung metastatic growth +/− dox via bioluminescence imaging. Barcode-qPCR was performed on lung genoms the relative abundance of each cell population in the lung metastatic burden. Each dot represents lung metastases from an individual animal.