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Figure 2 | Cell Communication and Signaling

Figure 2

From: Stoichiometry and intracellular fate of TRIM-containing TCR complexes

Figure 2

The stoichiometry of the TRIM-containing αβTCR. (A) Digitonin-solubilized scTCRs from 31.13scTCRβ cells were purified from NP-sepharose affinity columns using free NIP for elution. Before loading on a native gel, the TCR was incubated with the indicated amounts of the anti-TRIM antibody TRIM-7, and after separation, immunoblotted with anti-ζ. αβTCR complexes bound to one or two antibodies are marked with a or b, respectively. Ferritin was used as a marker protein as in figure 1. (B) αβTCRs were purified as in (A) and after separation by a 1D BN-PAGE, each lane was subjected to a 2D BN-PAGE into which the indicated amounts of the anti-CD3ε antibody UCHT1 had been loaded. The anti-ζ antiserum was used for Western blot detection. Arrows: small, the TCRαβ CD3ε2γδζ2 TCR; and large, the TCRαβ CD3ε2γδζ2TRIM2 TCR. (C) a schematic of the TRIM-containing large αβTCR complex is shown.

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