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. 1999 Aug;10(8):2493–2506. doi: 10.1091/mbc.10.8.2493

Figure 8.

Figure 8

Radiation-induced cleavage of 21-kDa pro-TGFα into a 5-kDa active TGFα fragment. EGFR-antisense cells were cultured as described in MATERIALS AND METHODS. Cells were irradiated (2 Gy), and 0, 30, 120, 180, and 300 min after irradiation (as indicated) the media were removed from these cells. Cells were not washed. Cells were immediately lysed in homogenization buffer, followed by immunoprecipitation using an antibody that recognizes both full-length pro-TGFα (∼21.5 kDa) and cleaved active TGFα (∼5 kDa) but not residual proteolytically cleaved TGFα forms (∼17–12 kDa). Proteins were resolved from immunoprecipitates by SDS-PAGE followed by immunoblotting using the same immunoprecipitating antibody, which recognizes pro-TGFα (∼21.5 kDa) and cleaved active TGFα (∼5 kDa), as indicated. A representative experiment is shown (n = 3). Exposure time, 4 min.