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. 2013 Oct 11;8(10):e75801. doi: 10.1371/journal.pone.0075801

Figure 3. p97 plays different roles in release of CTA1 and TCRα into the cytosol.

Figure 3

(A) CT-intoxicated cells transfected with vector, p97-His (WT p97) or p97-QQ-His (QQ p97) were subjected to the in vitro retro-translocation assay. P1, S1′ and S2′ were analyzed by immunoblotting with the indicated antibodies. (B) CE was incubated with a control antibody or antibody against p97. Immune-complexes were precipitated using protein A agarose beads and the resulting supernatants were analyzed by immunoblotting with the indicated antibodies. (C) P1 derived from CT-intoxicated cells was incubated with either mock- or p97-depleted CE. The resulting S2′ was analyzed by immunoblotting with the indicated antibodies. (D) Cells expressing TCRα-HA and co-transfected with vector, WT p97 or QQ p97 were treated with epoxomicin and subjected to the in vitro retro-translocation assay. The resulting P1, S1′ and S2′ were analyzed by immunoblotting with the indicated antibodies. (E) Same as in (C) except P1 was derived from epoxomicin-treated cells expressing TCRα-HA.