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Interaction of HlyA with cholesterol-depleted erythrocytes. Thirty micrograms of HlyA was incubated with 100 μl of cholesterol-depleted ghost erythrocytes for 30 min at 37 °C. Cells were lysed with 1% Triton X-100, and insoluble cell components were then separated by sucrose density gradient centrifugation. The gradient fractions were analyzed by immunoblotting with anti-HlyA (A) and anti-Flotillin-1 (B) antibodies.
