Figure 5
Figure 5. Confocal microscopy of fibrin clots show Aβ binding to fibrin fibrils. Fibrin clots were formed with or without Aβ42 as described in “Aβ binding to fibrin(ogen)” to determine the location of Aβ binding. (Top row) Fibrin visualized with Alexa Fluor 488–labeled fibrinogen (green); (bottom row) Aβ visualized with HiLyte Fluor 555–labeled Aβ (red); (A-B) control clot. (C-D) Clot with only unlabeled Aβ42 shows that fibrin fibers and irregular clusters do not produce signal in the red channel. (E-F) Clot formed with unlabeled Aβ42 and HiLyte Fluor 555–labeled Aβ42 shows colocalization between Aβ and fibrin fibers as well as Aβ and irregular clusters (arrowhead). (G-H) Clot formed without Alexa Fluor 488–labeled fibrinogen but with both unlabeled and labeled Aβ42 shows Aβ signal in the fibrin fiber pattern, confirming that Aβ signal is not Alexa Fluor 488 signal detected in the red channel. (I-J) Clot formed with unlabeled Aβ42 and HiLyte Fluor-555–labeled Aβ1-9 does not have Aβ signal along fibrin fibers or in aggregates, indicating that the Aβ42 signal represents specific Aβ-fibrin(ogen) binding and not fluorophore entrapment. Images are representative of ≥ 3 experiments.

Confocal microscopy of fibrin clots show Aβ binding to fibrin fibrils. Fibrin clots were formed with or without Aβ42 as described in “Aβ binding to fibrin(ogen)” to determine the location of Aβ binding. (Top row) Fibrin visualized with Alexa Fluor 488–labeled fibrinogen (green); (bottom row) Aβ visualized with HiLyte Fluor 555–labeled Aβ (red); (A-B) control clot. (C-D) Clot with only unlabeled Aβ42 shows that fibrin fibers and irregular clusters do not produce signal in the red channel. (E-F) Clot formed with unlabeled Aβ42 and HiLyte Fluor 555–labeled Aβ42 shows colocalization between Aβ and fibrin fibers as well as Aβ and irregular clusters (arrowhead). (G-H) Clot formed without Alexa Fluor 488–labeled fibrinogen but with both unlabeled and labeled Aβ42 shows Aβ signal in the fibrin fiber pattern, confirming that Aβ signal is not Alexa Fluor 488 signal detected in the red channel. (I-J) Clot formed with unlabeled Aβ42 and HiLyte Fluor-555–labeled Aβ1-9 does not have Aβ signal along fibrin fibers or in aggregates, indicating that the Aβ42 signal represents specific Aβ-fibrin(ogen) binding and not fluorophore entrapment. Images are representative of ≥ 3 experiments.

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