FIGURE 3.

LRP1 is involved in the regulation of TJ integrity in the apoE3-BBB model. A–D, apoE3- and apoE4-BBB models were cultured for 7 days. On day 7, the in vitro BBB models were treated with RAP (A) (1 μm) or the anti-LRP1 (B), anti-LDLR (C), or anti-VLDLR antibody (D) (25 μg/ml) for 4 h at 37 °C. After treatment, mBECs were harvested using a cell scraper, and the obtained cell lysates were subjected to Western blotting using anti-phosphorylated PKCη and anti-PKCη antibodies. E, apoE3- and apoE4-BBB models were cultured for 7 days. On day 7, the in vitro BBB models were treated with the anti-LRP1 antibody or isotype IgG (25 μg/ml) for 4 h at 37 °C. After treatment, mBEC lysates were immunoprecipitated with the polyclonal anti-Thr(P) antibody, and the immunoprecipitates were subjected to Western blotting using the anti-occludin antibody or anti-phosphooccludin antibody. F, apoE3- and apoE4-BBB models were cultured for 7 days and then treated with the anti-LRP1 antibody or isotype IgG (25 μg/ml) for 4 h at 37 °C. After treatment, TEER was measured, and the values are presented as Ω × cm². The data presented are means ± S.D. (error bars) (n = 3). *, p < 0.005 compared with the values of apoE3-BBB models treated with control IgG (Dunnett's test).