Fig. 3

HDL reduces Aβ42 uptake and LRP1 levels in SMC and blocking LRP1 with RAP reduces Aβ42 accumulation in bioengineered vessels. SMC were grown as 2D monolayers for 2 d before treating with 1 μM FITC-Aβ42 without or with 200 μg/mL of HDL for 3 h (a) or 24 h (b) before dissociating the cells and counting using flow cytometry. c SMC were grown in 2D culture for 3 d before treating with FITC-Aβ42 (5 μM, yellow) with 200 μg/mL of HDL or BSA for 24 h and imaged using microscopy. d SMC were grown for 3 days in 2D culture were treated with the indicated HDL concentration. LRP1 protein levels were measured by Western blot and normalized to GAPDH. e Aβ42 monomers (1 μM) were injected into the tissue chamber concomitantly with circulating 200 μg/mL of HDL or BSA through the lumen. After 24 h, tissues were lysed in RIPA. LRP1 protein levels were measured in bioengineered vessels using Western blot and normalized to GAPDH. f Bioengineered vessels were treated with 1 μM Aβ42 with or without recombinant RAP in the circulation media to block LRP1. After 24 h, tissues were lysed in RIPA and Aβ42 accumulation was measured by ELISA. Points in graphed data represent individual bioengineered vessels, bars represent mean, error bars represent ±SEM and analysed by Student’s t-test or one way ANOVA *P < 0.05