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. Author manuscript; available in PMC: 2014 Aug 20.
Published in final edited form as: Oncogene. 2013 Feb 18;33(8):986–995. doi: 10.1038/onc.2013.33

Figure 2. The engineered F-TrCP-Shc E3 ligase selectively targets the activated ErbB family members for degradation and depletes ErbB2 RTK on the cell surface.

Figure 2

SKBR3 (A) and BT474 (B) cells infected with the indicated recombinant retroviruses were lysed and 50 μg of each cell lysate was analyzed by SDS-PAGE and Western blotting with the indicated antibodies. C. SKBR3 and BT474 cells were plated in 6-well plates for 24 hours, trypsinized and washed in FACS buffer. The live cells were stained with anti-ErbB2 antibody (Santa Cruz) against the extracellular domain of ErbB2. Results are the mean for three replicate experiments. D. Dose-dependent degradation of endogenous ErbB2 upon infection of adenoviral F-TrCP-Shc was determined by Western blotting with the indicated antibodies. E. Depletion of ErbB2 on the cell membrane by adenoviral F-TrCP-Shc. ErbB2 (red) was detected by immunofluorescent staining with the anti-ErbB2 antibody (R&D Systems). Infected cells were marked by GFP expression from the adenoviral vector. The nuclei were visualized by DAPI staining (blue). White arrow, a low F-TrCP-Shc expressing SKBR3 cell.