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. Author manuscript; available in PMC: 2018 Jun 22.
Published in final edited form as: Cell. 2011 Mar 4;144(5):703–718. doi: 10.1016/j.cell.2011.02.003

Figure 1. A Genetic Screen for Tumor Suppressors Identifies PTPN12.

Figure 1

(A) Schematic of genetic screen for suppressors of HMEC transformation. A pool of retroviral shRNAs was transduced into TLM-HMECs in duplicate, and assessed for anchorage-independent proliferation. shRNAs were PCR amplified and sequenced from macroscopic colonies.

(B) Depletion of PTPN12. PTPN12 protein expression in TLM-HMECs transduced with vectors expressing the indicated shRNAs with quantification below.

(C) PTPN12 loss of function transforms TLM-HMECs. Anchorage-independent proliferation in TLM-HMECs transduced with the indicated shRNAs.

(D) Restoring PTPN12 expression suppresses transformation by PTPN12 shRNA. Anchorage-independent proliferation in TLM-HMECs transduced with control or PTPN12-shRNA in combination with PTPN12 cDNA as indicated.

(E) PTPN12 regulates acinar morphogenesis. MCF10A cells expressing the indicated shRNAs were analyzed for 3D acinar morphogenesis in vitro (day 15 after seeding) and quantified for the number of aberrant mammary acini.

(F) The enzymatic activity of PTPN12 is required for transformation suppression. TLM-HMECs expressing a PTPN12 shRNA were transduced with lentivirus-encoding control or shRNA-resistant PTPN12-C231S mutant cDNA and assessed for PTPN12 expression by western (top) and anchorage-independent proliferation (bottom).

Error bars represent standard error.