Figure 3
From: Expression, intracellular targeting and purification of HIV Nef variants in tobacco cells
Cytosolic Nef variants are predominantly soluble. Protoplasts transfected with the indicated plasmids were pulse labelled for 1 h and chased for 5 h. Cells were then homogenised in 12% (w/w) sucrose and homogenates were fractionated by centrifugation at 100,000 g for 30 min on a 17% sucrose pad. Total homogenates (T), membrane pellets (M) and soluble supernatant (S) fractions were then immunoprecipitated with anti BiP (upper panel) or anti FLAG (lower panel) antisera. Numbers at left indicate molecular weight markers in kDa.