FIGURE 10.

R-CRT is ubiquitinated. CHO cells were co-transfected with HA epitope-tagged Ub and Ub-R-CRT-EGFP or Ub-E1V-CRT-EGFP. Control cells were transfected with HA epitope-tagged Ub alone or with EGFP. Transfected cells were treated with MG132 for 18 h to cause accumulation of Ub conjugates, R-CRT-EGFP, or E1V-CRT-EGFP. Cell lysates were immunoprecipitated with anti-GFP Ab followed by immunoblotting (WB) with anti-GFP (A) and anti-HA (B) Abs. Lanes 3, 6, 9, and 12, immunoprecipitated proteins (IP); lanes 1, 4, 7, and 10, total cell lysate (Input): lanes 2, 4, 8, and 11, flow-through (Unbound).