Figure 3. PTPN12 Suppresses Transformation by Inhibiting HER2/EGFR Signaling.
(A) EGFR and HER2 depletion in TLM-HMECs. TLM-HMECs expressing control, EGFR, or HER2-targeting shRNAs were analyzed by western blotting for EGFR, HER2, and vinculin (loading control) as indicated.
(B) EGFR and HER2 RTKs are required for cellular transformation upon PTPN12 depletion. TLM-HMECs encoding an inducible PTPN12-shRNA were transduced with the indicated shRNAs and assessed for anchorage-independent proliferation ± dox.
(C) SHC depletion in HMECs. TLM-HMECs expressing control or SHC-targeting shRNAs were analyzed by western for SHC and vinculin as indicated.
(D) SHC is required for cellular transformation upon PTPN12 depletion. TLM-HMECs encoding an inducible PTPN12-shRNA were transduced with the indicated shRNAs and assessed for anchorage-independent proliferation ± dox.
(E) HER2/EGFR inhibitors block PTPN12 depletion-induced transformation. TLM-HMECs expressing the indicated shRNAs were assessed for anchorage-independent growth ± a HER2/EGFR inhibitor (lapatinib).
(F) Transformation by PTPN12 inactivation requires MAPK signaling. TLM-HMECs expressing the indicated shRNAs were assessed for anchorage-independent proliferation ± a MEK inhibitor (U0126).
Error bars represent standard error.
