Figure 4.

Degradation of UPS reporter protein CG*-GFP, is inhibited in yeast in the presence of FUS and overexpression of Sis1 mitigates this effect. Cells were grown in liquid plasmid selective 2% galactose medium for 48 hours to overexpress control (p2039) or FUS (p2049). In b and c GAL1-controlled Sis1 (p1767) or vector (p1768) were also expressed. Cycloheximide (0.5 or 0.7 mg/mL) was added, and cells were collected at the minutes (Min) indicated. Protein was extracted, and run on SDS-PAGE. The blots were probed with anti-GFP (1:10,000, Roche) to detect CG*-GFP levels (left) and anti-Pgk1 (1:10,000) as an internal loading control (right). Strains used were L1749 [PIN⁺] (a and b) and L2910 [pin^-] (c).