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. 2002 Nov 4;99(23):14728–14733. doi: 10.1073/pnas.232562799

Fig 3.

Fig 3.

Immunoblotting and autoradiography of gp130 and IL-6R. U266 cells (1 × 107 cells) were stimulated with or without 1 μg/ml rhIL-6 for 15 min in the presence or absence of MDL-A (30 or 100 μM, respectively). Cell lysates were immunoprecipitated with anti-IL-6R antibody, electrophoresed, and then blotted with anti-gp130 antibody (A) or anti-IL-6R antibody (B) for Western blot. In another experiment, cell lysate was also immunoprecipitated with anti-gp130 antibody, electrophoresed, and then blotted with anti-gp130 antibody (C) for Western blot. (A and B) Lanes 1–4 were incubated with 1.2 kBq [3H]MDL-A for autoradiography. (C) Lanes 5–7 were cut and incubated with 1.2 kBq [3H]MDL-A in the absence (lane 5) or presence of unlabeled MDL-A (30 μM, lane 6 or 100 μM, lane 7). a, Immunoblotting bands. b, Autoradiography in each group. Lane 1, no treatment; lane 2, 1 μg/ml rhIL-6; lane 3, 1 μg/ml rhIL-6 + 30 μM MDL-A; lane 4, 1 μg/ml rhIL-6 + 100 μM MDL-A.