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. 2007 Dec 19;82(5):2358–2366. doi: 10.1128/JVI.01931-07

FIG. 3.

FIG. 3.

Detection of TM trimers of activated Env by 2D BN-PAGE/nonreducing SDS-PAGE. Samples of [35S]Cys-labeled Mo-MLV were lysed in OG-EDTA HN buffer in the presence or absence of NEM for 20 min at 37°C to generate IAS Env and activated Env. A portion of the latter sample was subsequently cross-linked with DSP. Samples were subjected to 1D and 2D PAGE. (A and B) Phosphorimages of nonreducing SDS-PAGE and BN-PAGE of IAS Env (lanes 1) and activated Env (lanes 2). (C and D) 2D BN-PAGE/nonreducing SDS-PAGE of IAS Env (C) and activated Env (D). (E and F) 2D BN-PAGE/nonreducing SDS-PAGE of un-cross-linked (E) and cross-linked (F) activated Env. (C′) Cutout of the marked region of the second-dimension gel with SU-TM complexes at lower contrast. The oligomeric states of the complexes in the first dimension (BN-PAGE) are indicated. The marker lane (1D) in the 2D BN-PAGE/nonreducing SDS-PAGE contains the respective sample separated in nonreducing SDS-PAGE only. Viral proteins and their oligomeric state (C) in the first dimension (BN-PAGE) are indicated.