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. Author manuscript; available in PMC: 2021 Oct 11.
Published in final edited form as: Cell Rep. 2021 Sep 21;36(12):109717. doi: 10.1016/j.celrep.2021.109717

Figure 1. The cytosolic Hsp40 Ydj1 is required for the ERAD of SZ*.

Figure 1.

(A) The stability of SZ* from wild-type and ydj1Δ yeast expressing SZ* and the indicated vector in a CEN plasmid was determined in a cycloheximide chase. The QAA mutant lacks the Hsp70-interacting HPD motif. G6PD in this and other figures controls for equal loading. Data are means ± SE of 3–6 independent experiments; black asterisks apply to the +vector (ydj1Δ) versus +Ydj1 (ydj1Δ) comparison, and red asterisks apply to the +QAA mutant (ydj1Δ) versus +Ydj1 (ydj1Δ) comparison. **p < 0.005, ***p < 0.0005.

(B) HA-tagged SZ* was immunoprecipitated from wild-type yeast containing a vector control or ydj1Δ yeast expressing SZ* and Ydj1 or the QAA mutant. Cultures were shifted to 37°C or remained at 26°C for 1 h prior to harvest. A total of 1% of the input was analyzed to show equal loading. –LYS, indicates a control experiment performed in the absence of cell lysate. The asterisk denotes a contaminating species.

(C) This graph depicts relative Ydj1 binding to SZ* in yeast incubated at 26°C or 37°C, as quantified from the data in part (B). Data are the means ± SE of 3 independent experiments; *p < 0.05; NS, not significant.