Fig. 3. Solubility studies and time-course of expression of PfSec31p. (A) Mature-stage-parasitised erythrocytes (FAC8 strain, 10⁶ cells) were either sonicated in PBS (lanes a,b), or agitated in 2% Triton X-100 (lanes c,d) for 20 minutes at 4°C or sonicated in 50 mM NaHCO₃, pH 10 (lanes e,f) and the samples kept for 30 minutes at 4°C. The soluble and particulate fractions were separated by centrifugation and the supernatant (lanes b,d,f) and pellet (lanes a,c,e) fractions were subjected to SDS-PAGE (7.5% acrylamide) and transferred to a nitrocellulose membrane. The filters were incubated with antiserum against recombinant PfSec31(WD) followed by HRP-conjugated anti-rabbit IgG and visualised using ECL. (B) Aliquots of a synchronous culture of [³⁵S]-methionine/cysteine-labelled parasitised erythrocytes (FAC8 strain, 10% parasitemia) were harvested at the ring stage (lane a), the trophozoite stage (lane b), the schizont stage (lane c) or the ring stage of the second cycle (lane d). The samples were solubilised, immunoprecipitated with anti-PfSec31(WD) antiserum, subjected to SDS-PAGE (7.5% acrylamide) and visualised by phosphorimage analysis. A 160 kDa band that might correspond to a full-length monomeric form of PfSec31p is marked with an arrow.

Return to article