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

Figure 2. Proteasome degrades cytosol-localized deglycosylated TCRα but not CTA1.

Figure 2

(A) 293T cells intoxicated with or without CT were co-treated with or without epoxomicin for two hours before being subjected to the in vitro retro-translocation assay. P1, S1′ and S2′ were analyzed by immunoblotting with the indicated antibodies. (B) Cells transfected with vector or TCRα-HA were treated with or without epoxomicin for two hours before permeabilization to generate P1 and S1′. P1 from cells treated with or without epoxomicin were incubated with buffer, CE, or epoxomicin-treated CE to generate S2′. Fractions were analyzed by immunoblotting with the indicated antibodies. Observed TCRα-HA species were glycosylated to various degrees (+CHO). (C) Cells transfected with TCRα-HA were subjected to the in vitro retro-translocation assay to generate S2′. Where indicated, samples were treated with either Endo H or PNGase F at two different concentrations and then analyzed by immunoblotting with the indicated antibodies. Fully deglycoslyated species are denoted as (−CHO).