![Fig. 6. Transfer of hemolysis-inducing and glycophorin-cleaving activity. / E were treated with increasing concentrations of P1 (•), P2 (○), orL. intermedia venom (*), washed, and incubated with the same volume and number of calcein-AM loaded E or buffer. After 30 minutes of incubation at 37°C, cells were analyzed for autologous C lysis (A, B) or for the expression of GPC (C). (A) Total hemoglobin release induced by NHS determined spectrophotometrically at 541 nm. (B) Release of entrapped calcein determined fluorometrically. [C] E samples incubated with buffer or with P2 (2.5 and 5.0 μg/mL) were incubated with the same volume of untreated E (□) or with buffer (▪) for 30 minutes at 37 °C and analyzed by flow cytometry for expression of GPC using the monoclonal antibody Bric4. Results are expressed as the percentage of fluorescence of untreated E.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/95/2/10.1182_blood.v95.2.683/6/bloo00248006x.jpeg?Expires=1724272966&Signature=i37x3dJDA69~0TD~Vt-yCxISzD2OZ24qeIgp3FGMFKrVzY36iNnh5Ti6zsZiXdjFEFYAlsDtno5Mhkv5MwPu2JVt0fF3~WwBhzlm9iDPa-NsNbtocxgTUGcAjyp67YW~q4AmngZdcoP4hlOaAIxbVXBUekBDyF8l-7QtDgdigX4iqdW59usYaoyiyUvA51m1hyBvVtA5P5BGr2ix~JzxyG7qnqyMQGuwRb7ak~BTj4aPd4uCwV0XdMITVhXd53~YU~laCAQx957XR~u7Q5udK~lzHeWb9OENmSKVFbI-EFLxNu1pihg56ii7-2PF8W~Mt6l32dvMASkZXdbpvjRLVw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Transfer of hemolysis-inducing and glycophorin-cleaving activity.
E were treated with increasing concentrations of P1 (•), P2 (○), orL. intermedia venom (*), washed, and incubated with the same volume and number of calcein-AM loaded E or buffer. After 30 minutes of incubation at 37°C, cells were analyzed for autologous C lysis (A, B) or for the expression of GPC (C). (A) Total hemoglobin release induced by NHS determined spectrophotometrically at 541 nm. (B) Release of entrapped calcein determined fluorometrically. [C] E samples incubated with buffer or with P2 (2.5 and 5.0 μg/mL) were incubated with the same volume of untreated E (□) or with buffer (▪) for 30 minutes at 37 °C and analyzed by flow cytometry for expression of GPC using the monoclonal antibody Bric4. Results are expressed as the percentage of fluorescence of untreated E.
