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. 2004 Mar 1;164(5):769–779. doi: 10.1083/jcb.200307137

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Copyright © 2004, The Rockefeller University Press

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Figure 2. — Shedding of EGFR ligands in wild-type primary MEFs. (A) Diagram of a typical shedding experiment (see text for details). (B) Detection of shed AP-tagged EGFR ligands after renaturation in SDS gels (see Materials and methods for details). The left lane shows the AP-tagged forms of TGFα, amphiregulin, epiregulin, HB-EGF, betacellulin, and EGF released in 1 h into the supernatant of a single well each of transfected mEF under resting conditions. The next lane shows the EGFR ligands released in 1 h from the same well after addition of PMA, a phorbol ester that stimulates ectodomain shedding. The third lane shows EGFR ligands released from a separate well in 1 h under resting conditions, and the fourth lane shows the released EGFR ligands in that same well after addition of the hydroxamate-based metalloprotease inhibitor batimastat (BB94).

Figure 2. — Shedding of EGFR ligands in wild-type primary MEFs. (A) Diagram of a typical shedding experiment (see text for details). (B) Detection of shed AP-tagged EGFR ligands after renaturation in SDS gels (see Materials and methods for details). The left lane shows the AP-tagged forms of TGFα, amphiregulin, epiregulin, HB-EGF, betacellulin, and EGF released in 1 h into the supernatant of a single well each of transfected mEF under resting conditions. The next lane shows the EGFR ligands released in 1 h from the same well after addition of PMA, a phorbol ester that stimulates ectodomain shedding. The third lane shows EGFR ligands released from a separate well in 1 h under resting conditions, and the fourth lane shows the released EGFR ligands in that same well after addition of the hydroxamate-based metalloprotease inhibitor batimastat (BB94).