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. 2007 Dec 5;82(4):2004–2012. doi: 10.1128/JVI.02017-07

FIG. 6.

FIG. 6.

Protein A is tightly membrane associated in Δydj1 yeast. (A) Schematic of membrane flotation protocol. Whole-cell lysates were mixed with Nycodenz to 37.5%, loaded under a discontinuous 5 to 35% Nycodenz gradient, and spun to equilibrium at 100,000 × g. Equal-volume samples were taken from the top LD fraction and bottom HD fraction. LD fractions were further subjected to carbonate extraction at pH 11 for 30 min on ice and subsequently pelleted and separated into supernatant (CS) and pellet (CP) fractions by differential centrifugation. (B) Membrane flotation and carbonate extraction of membranes from wt, Δydj1, and complemented (Δydj1+ pYDJ1) yeast cells transformed with pFA-C/HA and pF1fs. Yeast PGK and porin are cytosolic and membrane proteins, respectively. (C) Membrane flotation and carbonate extraction of membranes from wt, Δydj1, and complemented (Δydj1+ pYDJ1) yeast cells transformed with pFA(HCV)-C/HA and pF1fs.