Figure 3.

The pool of ubiquitinated CPY^* at the ER membrane increases with time, whereas Sec61p loses contact to CPY^*. (A) The immunoprecipitation experiment was carried out as described in Figure 2A using Δprc1 and Δprc1Δder3 strains expressing either CPY^* or CPY^*HA₃ (input). The cells were pre-treated for 0, 15 and 30 min with CHX. The immunoblot analysis included the detection of ubiquitinated proteins. The stars indicate unspecific cross-reactions. (B) Ratio of the ubiquitin, Sec61p, Cdc48 and Der3p signals to the CPY^*HA₃ signals in immunoprecipitates of Δprc1+CPY^*HA₃ shown in (A) using the program ImageJ 1.41o.