The prefoldin complex stabilizes the von Hippel-Lindau protein against aggregation and degradation
Fig 2
A conditional expression system to produce GFP-pVHL213 in fission yeast.
A) Scheme of the integration cassette GFP-pVHL213 at the leu1 locus, GFP-pVHL213 is under the thiamine-repressible promoter nmt1. B) Reversed fluorescent (left) or phase contrast (right) images of leu1::VHL213 and leu1::GFP cells in the OFF (upper panels) or ON (lower panels) conditions. White and black arrows depict large and small aggregates, respectively. C) Western blot analysis of GFP-pVHL213 or GFP expression (ON condition) in leu1::VHL213 and leu1::GFP cells, respectively: Soluble (S) and Insoluble (I) fractions are shown. Cdc2 was used as a loading control. D) Aggregation of GFP-pVHL213 was compared between leu1::VHL213 cells transformed with empty control vectors (v1/v2) or v1-ElonginB and v2-Elongin C (+ELOB+ELOC) expressing plasmids. Left: reversed fluorescent images and right: histograms representing the percentage of large (upper panel) and small (lower panel) aggregate-containing cells (mean±s.d. from three independent experiments; **, p<0.01, Student t test). E) Western blot analysis of GFP-pVHL213 (VHL213), Elongin B (ELOB), Elongin C (ELOC) in cells: Soluble (S) and Insoluble (I) fractions are shown. Cdc2 was used as a loading control. Bars: 5 μm.