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. 2008 Apr 7;28(12):3932–3942. doi: 10.1128/MCB.02191-07

FIG. 1.

FIG. 1.

Steroid and thyroid hormones stimulate MED1 phosphorylation via activated MAPK-ERK signaling. (A to D) HeLa, HeLa-TR, MCF-7, and HeLa-AR cells were serum starved and then treated with EGF, U0126, T3, E2, or DHT (+) or untreated (−), as indicated (see Materials and Methods). The whole-cell lysate was then probed by immunoblotting for ERK1/2 expression (bottom) and ERK1/2 phosphorylation (top). (E to H) HeLa, HeLa-TR, MCF-7, and HeLa-AR cells were serum starved and then pulsed with 32Pi with or without hormones, as indicated. Parallel reactions lacking 32Pi were performed side by side. Native MED1 was immunoprecipitated with anti-MED1 antibodies and resolved by SDS-PAGE. Reactions containing 32Pi were detected by autoradiography (top), while reactions lacking 32Pi were detected by anti-MED1 immunoblotting (bottom).