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Figure 2


Fig. 2. Phosphoinositide binding of the A. aegypti ARH-like protein. (A) Human GST-ARH (60 µg/ml, residues 1-308; lanes a-d), GST-AaARH (residues 21-292; lanes e-h) or GST-AaARH PTB domain (residues 21-192, lanes i,j,k,l) were incubated with synthetic liposomes without (lanes a,b,e,f,i,j) or with PtdIns(4,5)P2 (lanes c,d,g,h,k,l) on ice in the presence of 100 µg/ml BSA. After centrifugation, aliquots of 3% of each supernatant (S) and 25% of each pellet (P) were resolved by SDS-PAGE and stained with Coomassie Blue. The position of the molecular mass standards (in kDa) is indicated. (B) GST-AaARH (60 µg/ml, residues 21-292, lanes a-f) or GST-ARH (residues 1-308, lanes g,h) were incubated with PtdIns(4,5)P2 liposomes in presence of 100 µg/ml BSA and either no (lanes a and b) or increasing concentrations (3 µg, lanes c,d or 6 µg, lanes e-j) of clathrin. After centrifugation, aliquots of 3% of each supernatant (S) and 25% of pellets (P) were resolved by SDS-PAGE and stained. The position of the clathrin heavy chain (HC) is indicated.





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